Capillarys / Minicap Urine

CAPILLARYS / MINICAP URINE
Ref. 2013
2012/05
CAPILLARYS / MINICAP URINE - 2012/05
INTENDED USE
The CAPILLARYS / MINICAP URINE kit is designed for the preparation of urine samples before separation of human urine proteins in alkaline buffer
(pH 10.0) with the CAPILLARYS and MINICAP Systems.
The CAPILLARYS and MINICAP perform automatically all sequences to obtain a urinary protein profile for qualitative analysis. The proteins, separated
in silica capillaries, are directly detected at an absorbance of 200 nm. The electrophoregrams can be interpreted visually to screen for any pattern
abnormalities.
For In Vitro Diagnostic Use.
PRINCIPLE OF THE TEST
Protein electrophoresis is a well established technique routinely used in clinical laboratories to screen samples for protein abnormalities (3, 4, 6, 12). The
CAPILLARYS and MINICAP have been developed to provide complete automation of this testing with fast separation and good resolution. In many
respects, the methodology can be considered as an intermediary type of technique between classical zone electrophoresis and liquid
chromatography (4, 14).
The CAPILLARYS and MINICAP Systems use the principle of capillary electrophoresis in free solution. With this technique, charged molecules are
separated by their electrophoretic mobility in an alkaline buffer with a specific pH. Separation also occurs according to the electrolyte pH and
electroosmotic flow (12).
The CAPILLARYS System has 8 capillaries functioning in parallel allowing 8 simultaneous analyses. The MINICAP System has 2 capillaries functioning
in parallel allowing 2 simultaneous analyses. The sample is prepared before the analysis by dialysis and concentration with buffer and then injected
by aspiration at the anodic end of the capillary. A high voltage protein separation is then performed and direct detection of the proteins is made at
200 nm at the cathodic end of the capillary. The capillaries are immediately washed with a Wash Solution and prepared for the next analysis with buffer.
Urinary proteins are separated at alkaline buffer into five different zones and detected in the following order : gamma zone, beta zone, alpha-2 zone,
alpha-1 zone and albumin with each zone containing one or more proteins.
REAGENTS SUPPLIED IN THE CAPILLARYS / MINICAP URINE KIT

WARNING : See the safety data sheets.
ITEM
PN 2013
Dialysis buffer (stock solution)
FOR OPTIMAL RESULTS :

All reagents from the same kit must be always used together and according to the package insert instructions.
PLEASE READ THE PACKAGE INSERT CAREFULLY.
1 vial, 480 mL
WARNING : Do not use marketed deionized water, such as water for ironing for example (risk of important capillaries damage). Use only
water with ultrapure quality, such as injection grade water.
DIALYSIS BUFFER
Preparation
The vial of the stock dialysis buffer should be diluted twice (v/v) with distilled or deionized water (1 volume of buffer + 1 volume of distilled or deionized
water).
After dilution, it contains : buffer solution pH 10.0 0.5 ; additives, non-hazardous at concentrations used, necessary for optimum performance.
Use
Dialysis buffer of urine samples for urinary proteins analysis in capillary electrophoresis (See "Samples preparation").
Storage, stability and signs of deterioration
Store the stock dialysis buffer at room temperature (15 to 30 C) or refrigerated (2 to 8 C). It is stable until the expiration date indicated on the kit
package or buffer vial labels. Avoid storage close to a window or to a heat source.
The diluted dialysis buffer is stable until the expiration date indicated on the kit package or buffer vial labels at room temperature or refrigerated.
NOTE : When dialysis buffer is stored between 2 to 8 C, it is recommended to allow reagent to come to room temperature prior to use.
DO NOT FREEZE.
Discard diluted buffer if it changes its appearance, e.g., becomes cloudy due to microbial contamination.
NOTE : During storage, stock buffer may turn yellow without any adverse effects on its performance.
- 19 -
SEBIA INSTRUCTIONS - English
CAPILLARYS URINE PROCEDURE

CAPILLARYS 1 & 2 release 5.4x
WARNING : The urine samples immunotyping can not be performed with this release.
REAGENTS REQUIRED (but not supplied with the CAPILLARYS / MINICAP URINE kit)
1. CAPILLARYS PROTEIN(E) 6 KIT (SEBIA, PN 2003)
Presentation, use, storage, stability and signs of deterioration

See the instruction sheets of the kit.
IMPORTANT : The components of CAPILLARYS PROTEIN(E) 6 kit are necessary to perform CAPILLARYS URINE procedure.
Uncoloured dilution segments supplied in the CAPILLARYS PROTEIN(E) 6 kit can not be used for the analysis of samples prepared with
Vivaspin 6 ml tubes.
2. DISTILLED OR DEIONIZED WATER
Use
For urine samples dilution, if necessary, and for the first dialysis (See "Samples preparation").
For rinsing capillaries in the SEBIA CAPILLARYS System, for capillary electrophoresis.
It is recommended to filter distilled or deionized water through 0.45 m filter before use.
To prevent microbial contamination, change the water every day. In case of longer storage, add 35 L/dL of ProClin 300.
IMPORTANT : Before filling the rinse container, it is recommended to wash it with plenty of distilled or deionized water.
3. CAPICLEAN
Composition
The vial of CAPICLEAN concentrated solution (SEBIA, PN 2058, 25 mL) contains : proteolytic enzymes, surfactants and additives nonhazardous at
concentrations used, necessary for optimum performances.
WARNING : See the safety data sheet.
Use
For weekly capillaries and sample probe cleaning in automated system CAPILLARYS, SEBIA, for capillary electrophoresis.
See the instruction sheets of CAPICLEAN, SEBIA.
IMPORTANT : Do not re-use the dilution segment after capillaries and probe cleaning.
Store CAPICLEAN refrigerated (2 8 C). It is stable until the expiration date indicated on the vial label. DO NOT FREEZE.
Precipitate or combined particles in suspension (floccules) may be observed in the CAPICLEAN vial without any adverse effects on its utilization.
Do not to dissolve this precipitate or these particles. It is recommended to collect only the supernatant.
4. SODIUM HYPOCHLORITE SOLUTION (for sample probe cleaning)
Preparation
Prepare a sodium hypochlorite solution (2 % to 3 % chloride) by diluting 250 mL 9.6 % chloride concentrated solution to 1 liter with cold distilled or
deionized water.
Use
For the sample probe cleaning in the CAPILLARYS System (weekly maintenance in order to eliminate adsorbed proteins from the probe).
See the SEBIA CAPILLARYS instruction manual.
Use the sample rack designed for the maintenance (No. 100).
Place a tube containing 2 mL (2 3 %) diluted chlorinated solution previously prepared, in position No. 1 on this sample rack.
Slide the sample rack No. 100 for maintenance in the CAPILLARYS System.
In the "MAINTENANCE" window which appears on the screen, select "Launch the probe cleaning (chlorinated sodium hypochlorite solution or CDT
wash solution)" and validate.
Store the working chlorinated solution at room temperature in a closed container, it is stable for 3 months. Avoid storage in sunlight, close to heat and
ignition source, and to acids and ammonia.
5. CAPILLARYS / MINICAP WASH SOLUTION
Preparation
Each vial of the stock CAPILLARYS / MINICAP Wash Solution (SEBIA, PN 2052, 2 vials, 75 mL) should be diluted up to 750 mL with distilled or
deionized water.
After dilution, the wash solution contains an alkaline solution pH 12.
Use
For washing the capillaries of CAPILLARYS. Additional wash solution is needed when the number of tests per series is below 40.
IMPORTANT : Before filling the wash solution container, it is recommended to wash the opening of the container, the connector and the tube with
plenty of distilled or deionized water to avoid salts deposit.
See instruction sheet for details.
Store the stock and working wash solutions in closed containers at room temperature or refrigerated. The stock wash solution is stable until the
expiration date indicated on the kit or wash solution vial label. Working wash solution is stable for 3 months.
Discard working wash solution if it changes its appearance, e.g., becomes cloudy due to microbial contamination.
- 20 -
NOTES :
The assays that were performed for the validation of reagents demonstrated that, for the different solutions and using an adapted equipment for the
reconstitution volume, a variation of 5 % on the final volume has no adverse effect on the analysis.
The distilled or deionized water used to reconstitute solutions, must be free of bacterial proliferation and mold (use a 0.22 m filter) and have a
resistivity higher than 10 Megohms x cm.
EQUIPMENT AND ACCESSORIES REQUIRED
1.
2.
3.
4.
CAPILLARYS System SEBIA : CAPILLARYS PN 1220 or CAPILLARYS 2 PN 1222.

Sample racks supplied with CAPILLARYS.
Container Kit supplied with CAPILLARYS : Rinse (to fill with distilled or deionized water), wash solution and waste containers.
For the dialysis and concentration of urine samples :
- SEBIA dialysis system, PN 9200 : 24 tubes of 20 mL with PES membrane 10 000 MWCO (10 KDa molecular weight cut-off),
or
- Dialysis and concentration system, type Vivaspin 6 ml, with PES membrane 10 000 MWCO (10 KDa molecular weight cut-off),
VIVASCIENCE,
or equivalent device giving the same performances and approved for clinical assays.
IMPORTANT : Please carefully read the instruction sheets of each dialysis and concentration device.
5. Swing bucket centrifuge with rotors for 17 mm diameter and 122 mm length tubes (when using Vivaspin 6 ml tubes) or for 30 mm diameter and
116 mm length tubes (when using SEBIA dialysis systems).
6. Green dilution segments SEBIA, PN 2081, for CAPILLARYS System, for the analysis of samples prepared with Vivaspin 6 ml tubes.
7. Pipettes : 200 L, 1 mL and 5 mL.
SAMPLES FOR ANALYSIS
Sample collection and storage
The analysis is carried out preferentially on fresh urine collected over 24 hours. Urine samples must be collected following established procedures
used in clinical laboratory testing.
NOTE : A urine sample collected over 24 hours is best suited for the analysis, but random urine sample can also be used for the qualitative analysis.
Samples may be stored for up to one week between 2 and 8 C.
For longer storage periods, it is recommended to keep samples frozen at 70 C (stable at least for one month).
IMPORTANT : Do not store samples at 20 C.
Thawed or improperly stored samples may show modified or additional fractions due to protein degradation.
NOTE : Urine samples should not be stored at room temperature!
Sample preparation
For the greatest recovery of urine proteins, it is highly recommended that centrifugal devices be used for the desalting / concentration
sample processing step of this assay.
IMPORTANT : The urine analysis technique by capillary electrophoresis requires 2 steps : a dialysis step and a concentration step of urine samples
with SEBIA dialysis systems (20 mL tubes) or with Vivaspin 6 ml tubes. Use only one tube per sample.
The following parameters are indicated for the use of SEBIA dialysis systems or Vivaspin 6 ml tubes with swing bucket centrifuge. Please
see carefully the instruction sheets of each dialysis and concentration device to use it with fixed angle centrifuge.
Before proceeding the analysis, prepare each sample according to one of the following procedures :
NOTE : Dilute twice (v/v) the stock dialysis buffer with distilled or deionized water to obtain the working dialysis buffer (1 volume of buffer + 1 volume
of distilled or deionized water).
Samples preparation with SEBIA dialysis systems (and only uncoloured dilution segments supplied in the CAPILLARYS PROTEIN(E) 6 kit)
1. Quantify total protein concentration in the urine sample to be analyzed.
2. Centrifuge 5 mL neat urine at 5 000 rpm for 10 minutes.
3. Collect the supernatant.
4. Prepare each sample in a new dialysis system according to its total protein concentration :
- When the total protein concentration is below 100 mg/dL : mix 1 mL urine with 9 mL distilled or deionized water in the tube.
- When the total protein concentration is between 100 and 300 mg/dL : mix 0.25 mL urine with 10 mL distilled or deionized water in the tube.
- When the total protein concentration is over 300 mg/dL : dilute first the urine to a protein concentration of 300 mg/dL with distilled or
deionized water ; then mix 0.25 mL diluted urine with 10 mL distilled or deionized water in the tube.
5. Homogenize the diluted urine.
6. Equilibrate the tubes with distilled or deionized water and centrifuge at 4 000 g* for 30 minutes.
WARNING : After concentration by centrifugation, the volume obtained must be up to 0.2 mL.
7. Add 10 mL working dialysis buffer per dialysis system.

8. Equilibrate the tubes with working dialysis buffer and centrifuge at 4 000 g* for 30 minutes.
9. After centrifugation, if the final volume is lower than 0.2 mL, add working dialysis buffer to a final volume of 0.2 mL (after the lower level of the
meniscus has equilibrated).
10. Homogenize the sample in the tube.
11. Take 200 L dialyzed urine and apply it directly in the well of a new uncoloured dilution segment.
12. Discard the dialysis system.
or
- 21 -

Samples preparation with Vivaspin 6 ml tubes (and only green dilution segments)
1. Quantify total protein concentration in the urine sample to be analyzed.
2. Centrifuge 5 mL neat urine at 5 000 rpm for 10 minutes.
3. Collect the supernatant.
4. Prepare each sample in a new Vivaspin 6 ml tube according to its total protein concentration :
- When the total protein concentration is below 100 mg/dL : mix 0.5 mL urine with 5.5 mL distilled or deionized water in the tube.
- When the total protein concentration is between 100 and 300 mg/dL : mix 0.125 mL urine with 6 mL distilled or deionized water in the tube.
deionized water ; then mix 0.125 mL diluted urine with 6 mL distilled or deionized water in the tube.
7.
8.
9.
10.
Discard the solution contained in the lower part of the Vivaspin tube.
Add 6 mL working dialysis buffer per Vivaspin tube.
Equilibrate the tubes with working dialysis buffer and centrifuge at 3 500 g* for 30 minutes.
After centrifugation, if the final volume is lower than 0.1 mL, add working dialysis buffer to a final volume of 0.1 mL (after the lower level of the
meniscus has equilibrated).
12. Take 100 L dialyzed urine and apply it directly in the well of a new green dilution segment.
13. Discard the Vivaspin tube.
IMPORTANT : Analyze the urine samples within a maximum of one day after their preparation. In order to limit protein adsorption onto the membrane
of the dialysis and concentration device (Vivaspin 6 ml tube or SEBIA dialysis system), it is not recommended to store the sample in the tube after
centrifugation but in a microtube stored refrigerated (between 2 and 8 C).
* See the centrifuge instruction manual to calculate the rpm number corresponding to the rotor.
The instructions for the use of dialysis and concentration devices with swing bucket centrifuge and fixed angle centrifuge are different, carefully check
the corresponding instruction sheets.
Samples to avoid
Avoid aged, improperly stored urine samples, fractions would be modified due to denaturation.
It is advised to observe the urine sample features after the first centrifugation (5 000 rpm for 10 minutes) (e.g., signs of hemolysis or turbidity).
Do not store samples in dialysis and concentration devices, some proteins may bind to the membrane.
Do not store more than 30 minutes dialyzed urine samples in dilution segment.
PROCEDURE
The CAPILLARYS system is a multiparameter instrument for urinary protein analysis on 8 parallel capillaries in the following sequence :
Bar code reading of samples-racks ;
Capillary washing ;
Injection of samples ;
Protein analysis and direct detection on capillaries.
The manual steps include :

Samples preparation (dialysis and concentration) ;
Samples application in dilution segment wells (uncoloured or green dilution segment according to the dialysis and concentration device used) ;
Placement of dilution segments with samples in sample-racks and samples identification ;
Placement of racks on the CAPILLARYS instrument ;
Removal of sample-racks after analysis.
PLEASE CAREFULLY READ THE CAPILLARYS INSTRUCTION MANUAL.
I. PREPARATION OF ELECTROPHORETIC ANALYSIS
1. Switch on CAPILLARYS instrument and computer.

2. Launch the software, the instrument automatically starts.
3. Analyse the urine samples prepared with the CAPILLARYS URINE kit with "URINE" analysis program from the CAPILLARYS instrument and
the CAPILLARYS PROTEIN(E) 6 buffer. To select "URINE" analysis program and place the CAPILLARYS PROTEIN(E) 6 buffer vial in the
instrument, please read carefully the CAPILLARYS instruction manual.
4. Load a sample rack with a new dilution segment filled with dialyzed urine samples : 100 L sample per well of a green dilution segment or
200 L sample per well of an uncoloured dilution segment. The sample rack will be ejected if the segment is missing.
IMPORTANT : If the number of samples to analyze is lower than 8, complete the dilution segment with distilled or deionized water.
5. Slide the complete sample carrier(s) into the CAPILLARYS system through the opening in the middle of the instrument. Up to 4 sample racks
can be introduced successively and continuously into the system.
6. Remove analyzed sample racks from the plate on the left side of the instrument.
7. Take off carefully used dilution segments from the sample rack and discard them.
WARNING : Dilution segments with biological samples have to be handled with care.
- 22 -
DILUTION - MIGRATION - DESCRIPTION OF THE AUTOMATED STEPS

1.
2.
3.
4.
5.
Bar codes are read on sample racks containing dilution segments and urine samples to analyze.
Capillaries are washed.
Samples are injected into capillaries.
Migration is carried out under constant voltage, controlled by Peltier effect, for about 4 minutes.
Proteins are detected directly by scanning at 200 nm and an electrophoretic profile appears on the screen of the system.
NOTE : These steps are described for the first introduced sample rack. The electrophoretic patterns appear after 10 minutes. For the following sample
rack, the first step (bar code reading) is made during analysis of the previous sample rack.
II. RESULT ANALYSIS
At the end of the analysis, the electrophoretic profiles can be analyzed and interpreted visually for pattern abnormalities. The different protein zone
positions (Albumin, Alpha 1, Alpha 2, Beta and Gamma) are identified on the screen and on the result report.
NOTE : For each of the 3 sample preparation procedures, the extent of the pattern visualized on the y-axis of the screen takes into account the total
protein concentration in each analyzed sample.
III. END OF ANALYSIS SEQUENCE
At the end of each analysis sequence, the operator must initiate the "stand by" or "shut down" procedure of the CAPILLARYS System in order to store
capillaries in optimal conditions.
IV. FILLING OF REAGENT CONTAINERS
The CAPILLARYS System has a reagent automatic control.

IMPORTANT : Please refer to the instructions for replacement of reagent containers respecting colour code for vials and connectors.
A message will be displayed when it is necessary to perform one of the following tasks :
Place a new buffer vial and / or ;
Fill the container with working wash solution and / or ;
Fill the container with filtered distilled or deionized water for rinsing capillaries and / or ;
Empty the waste container.
RESULTS
Interpretation
Interpretation is qualitative.
As an aid for patterns interpretation, it is possible to overlay the Normal Control Serum (SEBIA, PN 4785) to obtain a reference pattern. Reconstitute
the lyophilized serum with distilled or deionized water, dilute it 40 times in working dialysis buffer and analyze directly the diluted serum as a urine
sample.
Physiological proteinuria (1, 2, 7, 8, 9, 13) : It is weak, generally lower than 120 mg / 24 hours and there is no significant difference between electrophoretic
profiles from male and female. The major protein is albumin associated with traces of transferrin and immunoglobulins. A proteinuria higher than
120 mg / 24 hours must be considered as pathological.
A positive proteinuria (> 120 mg / 24 hours) should be followed up by a qualitative analysis of the eliminated proteins (5).
Urine electrophoresis is performed in order to detect monoclonal components, particularly a Bence Jones protein (free light chain), and other urinary
proteins. When compared to a normal pattern, an additional fraction, especially in the gammaglobulin or betaglobulin zone is related to a disorder (9, 10, 11).
Urine electrophoresis is efficient to detect monoclonal components (monoclonal immunoglobulins or free light chains) with low concentrations at about
2.0 mg/dL.
An identification by immunofixation with HYDRAGEL BENCE JONES, HYDRAGEL IF and HYDRAGEL URINE PROFIL(E) SEBIA kits and specific
antisera anti-free kappa and lambda light chains is recommended to characterize monoclonal or oligoclonal components.
Complementary tests, such as electrophoresis with SEBIA HYDRAGEL PROTEINURIE or HYDRAGEL URINE PROFIL(E) kits, or nephelometric
immunoassays, are necessary to characterize suspected tubular, glomerular or mixed proteinurias after capillary electrophoresis (3).
As an aid in interpretation of urinary protein electrophoregrams, see BIBLIOGRAPHY.
Particular cases :
- Sample urines with total protein concentration below 15 mg/dL (before preparation for analysis) may show a protein background (smear) on the
whole pattern.
- It is recommended to perform further analysis for Bence Jones protein when an abnormality is suspected in the Beta or Gamma zone whatever the
total protein concentration is.
- 23 -
Interference and Limitations
See SAMPLES FOR ANALYSIS.

Analyze only samples prepared with dialysis and concentration devices, see EQUIPMENT AND ACCESSORIES REQUIRED, such as Vivaspin 6 mL
tubes, SEBIA dialysis systems or equivalent device giving the same performances and approved for clinical assays.
A deficient sample dialysis may lead to non proteic residual fractions. When an interferent fraction is suspected, it is recommended to dialyse the urine
sample again.
Interferences such as drug or salts are eliminated during the dialysis step. If this step is not carefully followed, artifactual peaks might be observed.
Due to the resolution and sensitivity limits of zone electrophoresis, it is possible that some urinary proteins may not be detected with this method.
Troubleshooting
Call SEBIA when the test fails to perform even though when the instructions for the preparation, storage of materials, and for the procedure were
carefully followed.
Kit reagent Safety Data Sheets and information on elimination of waste products are available from SEBIA.
- 24 -
CAPILLARYS 1 & 2 release 6.x
1. CAPILLARYS PROTEIN(E) 6 KIT (SEBIA, PN 2003)

IMPORTANT : The components of CAPILLARYS PROTEIN(E) 6 kit are necessary to perform CAPILLARYS URINE procedure.
WARNING : Do not use uncoloured dilution segments supplied in the CAPILLARYS PROTEIN(E) 6 kit for the analysis of samples with
CAPILLARYS URINE technique.
2. NORMAL CONTROL SERUM
Composition
The Normal Control Serum (SEBIA, PN 4785) is obtained from a pool of normal human sera. The Normal Control Serum is in a stabilized lyophilised
form.
Intended Use
The Normal Control Serum is designed, before starting a new analysis sequence, for the normalization of capillaries and for the migration control
of human urinary proteins with CAPILLARYS URINE electrophoresis procedure.
Reconstitute each Normal Serum Control vial with 1 mL of distilled or deionized water. Allow to stand for 30 minutes and mix gently (avoid formation
of foam).
The reconstituted Normal Control Serum should be used as a normal human serum.
See the paragraph "Preparation of electrophoretic analysis" for the normalization of capillaries.
Before reconstitution, store the lyophilised Normal Control Serum refrigerated (2 to 8 C). It is stable until the expiration date indicated on the box or
vial labels.
Store the reconstituted Normal Control Serum at 2 8 C. Due to the risk of microbial contamination and denaturation, use it within one week.
The reconstituted serum may also be frozen (in aliquots) and stored at - 20 C for 2 months maximum.
Before use, store the thawed Normal Control Serum at 2 8 C and use it within the day.
NOTE : For optimal use with the CAPILLARYS system, it is recommended to split the Normal Control Serum into aliquots in microtubes before
freezing.
NOTE : During transportation, the lyophilized Control Serum can be kept without refrigeration (15 to 30 C) for 15 days without any adverse effects
on performance.
No test method can provide an absolute assurance of the absence of HIV, hepatitis B and C or other infectious agents. Therefore, handle the Control
Serum as a hazardous biological material.
This lot of Control Serum was found negative on assays approved by FDA or EU equivalent regulatory agency :
- against hepatitis B surface antigen ;
- for antibody to HCV ;
- for antibody to HIV1 and HIV2.
Use
For rinsing capillaries in the SEBIA CAPILLARYS System, for capillary electrophoresis.
4. CAPICLEAN
Composition
Use
For weekly capillaries and sample probe cleaning in automated system CAPILLARYS, SEBIA, for capillary electrophoresis.
IMPORTANT : Do not re-use the dilution segment after capillaries and probe cleaning.
- 25 -
Preparation
deionized water.
Use
For the sample probe cleaning in the CAPILLARYS System (weekly maintenance in order to eliminate adsorbed proteins from the probe).
See the SEBIA CAPILLARYS instruction manual.
Use the sample rack designed for the maintenance (No. 100).
Place a tube containing 2 mL (2 3 %) diluted chlorinated solution previously prepared, in position No. 1 on this sample rack.
Slide the sample rack No. 100 for maintenance in the CAPILLARYS System.
In the "MAINTENANCE" window which appears on the screen, select "Launch the probe cleaning (chlorinated sodium hypochlorite solution or CDT
wash solution)" and validate.
Preparation
deionized water.
Use
For washing the capillaries of CAPILLARYS. Additional wash solution is needed when the number of tests per series is below 40.
See instruction sheet for details.
NOTES :
1.
2.
3.
4.
5.
6.
7.
8.
9.
CAPILLARYS System SEBIA : CAPILLARYS PN 1220 or CAPILLARYS 2 PN 1222.

Sample racks supplied with CAPILLARYS.
Container Kit supplied with CAPILLARYS : Rinse (to fill with distilled or deionized water), wash solution and waste containers.
SEBIA dialysis system, PN 9200, 24 tubes of 20 mL with PES membrane 10 000 MWCO (10 KDa molecular weight cut-off) for the dialysis and
concentration of urine samples,
or equivalent device giving the same performances and approved for clinical assays which leads to a final volume of dialyzed and concentrated
urine sample of 500 L (volume of sample necessary to perform analyses with CAPILLARYS URINE and IMMUNOTYPING URINE techniques).
Swing bucket centrifuge with rotors for 30 mm diameter and 116 mm length tubes when using SEBIA dialysis systems.
Green dilution segments SEBIA, PN 2081, for CAPILLARYS System.
Pipettes : 200 L, 1 mL and 5 mL.
1.5 mL microtubes.
Hemolysing tubes (75 mm high and 13 mm in diameter).
- 26 -
Sample preparation (with SEBIA dialysis systems)
with SEBIA dialysis systems (20 mL tubes). Use only one tube per sample.
The following parameters are indicated for the use of SEBIA dialysis systems with swing bucket centrifuge. Please see carefully the
instruction sheets of each dialysis and concentration device to use it with fixed angle centrifuge.
Before proceeding the analysis, prepare each sample according to the following procedure :
Quantify total protein concentration in the urine sample to be analyzed.

Centrifuge 5 mL neat urine at 5 000 rpm for 10 minutes.
Collect the supernatant.
Prepare each sample in a new dialysis system according to its total protein concentration :
- When the total protein concentration is between 100 and 300 mg/dL : mix 0.5 mL urine with 19.5 mL distilled or deionized water in the tube.
deionized water ; then mix 0.5 mL diluted urine with 19.5 mL distilled or deionized water in the tube.
1.
2.
3.
4.
7. For dialysis, add 20 mL working dialysis buffer per dialysis system.

9. After centrifugation, a part of the sample may be left on the upper flat zone of the dialysis system (see the enclosed figure). With a micropipette,
collect this sample and transfer it in the lower well of the dialysis system. If the final volume is less than 0.5 mL, add working dialysis buffer to a
final volume of 0.5 mL (after the lower level of the meniscus has equilibrated).
0.75
0.5
0.2

11. Collect the dialyzed urine remaining in the tube and apply it in a 1.5 mL microtube.
of the dialysis and concentration device (SEBIA dialysis system), it is not recommended to store the sample in the dialysis system after centrifugation
but in a microtube stored refrigerated (between 2 and 8 C).
The instructions for the use of dialysis and concentration devices with swing bucket centrifuge and fixed angle centrifuge are different. The parameters
described above are indicated for the use of SEBIA dialysis systems with swing bucket centrifuge. Please see carefully the instruction sheets of each
dialysis and concentration device to use it with fixed angle centrifuge.
Samples to avoid
Do not re-use the urine sample applied in the dilution segment.
- 27 -
PROCEDURE
The CAPILLARYS system is a multiparameter instrument for urinary protein analysis on 8 parallel capillaries in the following sequence :
Bar code reading of samples-racks and samples to analyze ;
Samples application in dilution segment wells from microtubes containing the dialyzed samples ;
Capillary washing ;

Sample preparation (dialysis and concentration) ;
Placement of microtubes (without caps) containing the samples to analyze on holding hemolysing tubes in sample-racks ; each tube being identified
with the specific sample identification bar code label corresponding to the sample to analyze.
Placement of new 100 L green dilution segments in sample-racks ;
Placement of racks on the CAPILLARYS instrument ;
Removal after analysis of sample-racks and microtubes for potential analysis by immunotyping.
1. Switch on CAPILLARYS instrument and computer.

2. Launch the software, the instrument automatically starts.
3. The CAPILLARYS instrument must be used with "URINE" analysis program and the CAPILLARYS PROTEIN(E) 6 buffer. To select "URINE"
analysis program and place the CAPILLARYS PROTEIN(E) 6 buffer vial in the instrument, please read carefully the CAPILLARYS instruction
manual.
4. Normalize the capillaries with the Normal Control Serum (see the paragraph "Reagents required but not supplied").
IMPORTANT : It is necessary to normalize the capillaries before urine samples analysis in the CAPILLARYS instrument in order to analyze
them afterwards with CAPILLARYS IMMUNOTYPING URINE procedure.
- Apply 25 L reconstituted Normal Control Serum in a hemolysing tube and 2 mL of working dialysis buffer ; then, homogenize.
- Place this tube in position No. 1 on the sample rack No. 0 of the CAPILLARYS instrument intended for control sample, and a new green
dilution segment. A message will be displayed when the segment is missing.
- Start the analysis : Slide the sample rack No. 0 into the CAPILLARYS system, select "Normal Control" in the window which appears on the
screen and validate.
The results are then automatically considered by the software for the data analysis.
5. Analyse the urine samples prepared with the CAPILLARYS / MINICAP URINE kit :
- The sample rack contains eight positions for tubes. Place eight empty hemolysing tubes (used as holders) on each sample rack, and then,
the microtubes containing the dialyzed urines samples. Cut the cap of each microtube before using it.
- Keep the cap of each microtube for further storage of samples, if necessary.
IMPORTANT : It is recommended to identify each hemolysing tube holding the microtube which contains samples to analyze, with the specific
sample identification bar code label corresponding to the sample. This identification allows to determine the necessary dilution program to use
when performing the CAPILLARYS IMMUNOTYPING URINE technique.
- The bar code of each tube must be visible in the openings of the sample rack.
NOTE : If the number of samples to analyze is less than 8, complete the sample rack with tubes containing distilled or deionized water.
- Place a new green dilution segment on each sample rack. The sample rack will be ejected if the segment is missing.
- Slide the complete sample carrier(s) into the CAPILLARYS system through the opening in the middle of the instrument. Up to 13 sample
racks can be introduced successively and continuously into the system.
6. Remove analyzed sample racks from the plate on the left side of the instrument.
7. Take off carefully used dilution segments from the sample rack and discard them.
WARNING : Dilution segments with biological samples have to be handled with care.
8. Take off microtubes containing analyzed urine samples from the sample rack and store them refrigerated (between 2 and 8 C) for further
analysis with CAPILLARYS IMMUNOTYPING URINE technique, if necessary.
1.
2.
3.
4.
5.
6.
Bar codes are read on sample racks and on tubes with urine samples to analyze.
Samples are applied in dilution segment and the sample probe is rinsed after each sample.
NOTE : These steps are described for the first introduced sample rack. The electrophoretic patterns appear after 10 minutes. For the following sample
rack, the steps 1 and 2 (bar codes reading and sample application) are made during the step 5 of the previous sample rack.
II. RESULT ANALYSIS
NOTE : For each of the 3 sample preparation procedures, the extent of the pattern visualized on the y-axis on the screen takes into account the total
- 28 -

Abnormal fractions must be integrated using the "quantify peaks" tool of the software, then, the "Ratio urine", defined by the proportion of the abnormal
fraction area related to the Normal Control Serum proteins total area, is automatically calculated ; it will be displayed in the modify curves window.
This ratio will be used in the CAPILLARYS IMMUNOTYPING URINE technique to determine the sample dilution program to use ("HYPOGAMMA",
"STANDARD" or "HYPERGAMMA").
At the end of each analysis sequence, the operator must initiate the "stand by" or "shut down" procedure of the CAPILLARYS System in order to store
The CAPILLARYS System has a reagent automatic control.

IMPORTANT : Please refer to the instructions for replacement of reagent containers respecting colour code for vials and connectors.
RESULTS
Interpretation
As an aid for patterns interpretation, it is recommended to overlay a pattern obtained after the normalization of capillaries with the Normal Control
Serum to obtain a reference pattern.
2.0 mg/dL.
An identification is recommended to characterize monoclonal or oligoclonal components :
- by immunotyping with SEBIA CAPILLARYS IMMUNOTYPING kit or,
- by immunofixation with SEBIA HYDRAGEL BENCE JONES, HYDRAGEL IF and HYDRAGEL URINE PROFIL(E) kits and specific antisera anti-free
kappa and lambda light chains.
Particular cases :
whole pattern.

Analyze only samples prepared with dialysis and concentration devices, indicated in the paragraph "EQUIPMENT AND ACCESSORIES REQUIRED",
such as SEBIA dialysis systems or equivalent device giving the same performances and approved for clinical assays.
sample again.
Troubleshooting
carefully followed.
- 29 -
PERFORMANCE DATA
Standard materials, sample preparation and procedures were used. All electrophoregrams were evaluated visually.
Reproducibility within run
Reproducibility within run was demonstrated on two different pathological urine samples (one urine sample containing five protein fractions and one
sample with a Bence Jones protein) and on one normal sample (physiological urine). Each sample was prepared with dialysis buffer from two different
lots, then applied in all wells of dilution segments and analyzed using CAPILLARYS URINE procedure with the same analysis buffer.
All repeats gave concordant results within run and within the two dialysis buffer lots. Patterns corresponded to the type of each tested sample.
With CAPILLARYS URINE procedure, the urinary proteins were correctly identified in the two pathological samples, there were no false positives and
no differences were observed among the repeats. No protein was detected with the normal sample.
Reproducibility between runs and lot-to-lot
Reproducibility between runs was demonstrated on six different pathological urine samples containing one to seven protein fractions and on two
normal samples (physiological urines) prepared each with dialysis buffer from three different lots.
These samples were analyzed 10 times using CAPILLARYS URINE procedure for each dialysis buffer lot and with the same analysis buffer.
All repeats gave concordant results between runs and lot-to-lot.
With CAPILLARYS URINE procedure, the urinary proteins were correctly identified in the six pathological samples, there were no false positives and
no differences were observed among the repeats. For each urine sample tested, typical pattern and specific urinary proteins were detected as
expected for each sample type. No abnormal protein was detected with the normal samples.
Concordance Study
Concordance study was performed on 100 urine samples between CAPILLARYS URINE procedure and a commercially available agarose gel
electrophoresis system for urine analysis (without any urine concentration) : 19 normal urine samples and 81 different pathological urines have been
run on both techniques ; the total protein concentrations were comprised between 0.07 and 19.83 g/L.
This study demonstrated a 100 % agreement between the two techniques :
For the 19 normal urine samples : complete agreement (concordance).
For the 81 pathological urine samples : complete agreement (concordance).
For all the analyzed pathological urine samples, the results obtained with the individual procedures were in agreement and the urinary protein fractions
were detected on all pathological samples with each system.
Sensitivity
Serial dilutions were prepared with one pathological urine sample containing a Bence Jones protein (at concentration about 0.34 g/L) diluted in normal
urine and analyzed using the CAPILLARYS URINE procedure.
The minimal detection limit of the monoclonal component was about 2.0 mg/dL.
NOTE : According to the position of the monoclonal component and polyclonal background in the gamma and beta zones, the detection limit may vary.
- 30 -
MINICAP URINE PROCEDURE
1. MINICAP PROTEIN(E) 6 KIT (SEBIA, PN 2203)

IMPORTANT : The components of MINICAP PROTEIN(E) 6 kit are necessary to perform MINICAP URINE procedure.
2. NORMAL CONTROL SERUM
Composition
The Normal Control Serum (SEBIA, PN 4785) is obtained from a pool of normal human sera. The Normal Control Serum is in a stabilized lyophilised
form.
Intended Use
The Normal Control Serum is designed, before starting a new analysis sequence, for the normalization of capillaries and for the migration control
of human urinary proteins with MINICAP URINE electrophoresis procedure.
Reconstitute each Normal Serum Control vial with 1 mL of distilled or deionized water. Allow to stand for 30 minutes and mix gently (avoid formation
of foam).
The reconstituted Normal Control Serum should be used as a normal human serum.
See the paragraph "Preparation of electrophoretic analysis" for the normalization of capillaries.
Before reconstitution, store the lyophilised Normal Control Serum refrigerated (2 to 8 C). It is stable until the expiration date indicated on the box or
vial labels.
Store the reconstituted Normal Control Serum at 2 8 C. Due to the risk of microbial contamination and denaturation, use it within one week.
The reconstituted serum may also be frozen (in aliquots) and stored at - 20 C for 2 months maximum.
Before use, store the thawed Normal Control Serum at 2 8 C and use it within the day.
NOTE : For optimal use with the MINICAP system, it is recommended to split the Normal Control Serum into aliquots in microtubes before freezing.
NOTE : During transportation, the lyophilized Control Serum can be kept without refrigeration (15 to 30 C) for 15 days without any adverse effects
on performance.
No test method can provide an absolute assurance of the absence of HIV, hepatitis B and C or other infectious agents. Therefore, handle the Control
Serum as a hazardous biological material.
This lot of Control Serum was found negative on assays approved by FDA or EU equivalent regulatory agency :
- against hepatitis B surface antigen ;
- for antibody to HCV ;
- for antibody to HIV1 and HIV2.
Use
For rinsing capillaries in the SEBIA MINICAP System, for capillary electrophoresis.
4. CAPICLEAN
Composition
Use
For weekly sample probe cleaning in automated system MINICAP, SEBIA, for capillary electrophoresis.
IMPORTANT : For optimal use of the CAPICLEAN solution with the MINICAP system, it is necessary to use one bar code label intended to identify
the hemolysing tube holding the microtube which contains the diluted CAPICLEAN solution (cut the cap of the microtube before using it).
For later use, store the tube containing the diluted solution at 2 8 C. It must be used within the day.
Preparation
deionized water.
- 31 -
Use
For the sample probe cleaning in the MINICAP system (weekly maintenance in order to eliminate adsorbed proteins from the probe).
See the SEBIA MINICAP instruction manual.
Apply in a microtube 500 L of diluted chlorinated solution previously prepared.
Cut the cap of the microtube.
Place the microtube, located on a new hemolysing tube used as a support (identified with one bar code label specific to the sodium hypochlorite
solution) into position No. 1 on the rotating sampler of MINICAP.
Position a new reagent cup on the automated loading system for cups of MINICAP (a message will be displayed if the reagent cup is missing).
Slide the rotating sampler into the MINICAP system.
Close the doors of the MINICAP system, the cleaning sequence automatically starts.
IMPORTANT : For optimal use of the sodium hypochlorite solution with the MINICAP system, it is necessary to use one bar code label intended to
identify the hemolysing tube holding the microtube which contains the solution (cut the cap of the microtube before using it).
Preparation
deionized water.
For MINICAP, it is convenient to dilute only 25 mL of the stock solution to 250 mL with distilled or deionized water.
Use
For washing the capillaries of MINICAP.

NOTES :
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
MINICAP System, SEBIA, PN 1230.

Rotating sampler supplied with MINICAP.
Container Kit supplied with MINICAP : Rinse (to fill with distilled or deionized water) and waste container.
MINICAP Reagent cups, SEBIA (250 units), PN 2280.
Lids for bins for used reagent cups, SEBIA (12 units), PN 2286 : lids to close the bins containing used cups.
SEBIA dialysis system, PN 9200, 24 tubes of 20 mL with PES membrane 10 000 MWCO (10 KDa molecular weight cut-off) for the dialysis and
concentration of urine samples,
or equivalent device giving the same performances and approved for clinical assays which leads to a final volume of dialyzed and concentrated
urine sample of 500 L (volume of sample necessary to perform analyses with MINICAP URINE and MINICAP IMMUNOTYPING URINE
techniques).
Swing bucket centrifuge with rotors for 30 mm diameter and 116 mm length tubes when using SEBIA dialysis systems.
Pipettes : 200 L, 1 mL and 5 mL.
1.5 mL microtubes.
Hemolysing tubes (75 mm high and 13 mm in diameter).
- 32 -
Sample preparation (with SEBIA dialysis systems)
with SEBIA dialysis systems (20 mL tubes). Use only one tube per sample.
The following parameters are indicated for the use of SEBIA dialysis systems with swing bucket centrifuge. Please see carefully the
instruction sheets of each dialysis and concentration device to use it with fixed angle centrifuge.
Before proceeding the analysis, prepare each sample according to the following procedure :
Quantify total protein concentration in the urine sample to be analyzed.

Centrifuge 5 mL neat urine at 5 000 rpm for 10 minutes.
Collect the supernatant.
Prepare each sample in a new dialysis system according to its total protein concentration :
- When the total protein concentration is between 100 and 300 mg/dL : mix 0.5 mL urine with 19.5 mL distilled or deionized water in the tube.
deionized water ; then mix 0.5 mL diluted urine with 19.5 mL distilled or deionized water in the tube.
1.
2.
3.
4.
7. For dialysis, add 20 mL working dialysis buffer per dialysis system.

9. After centrifugation, a part of the sample may be left on the upper flat zone of the dialysis system (see the enclosed figure). With a micropipette,
collect this sample and transfer it in the lower well of the dialysis system. If the final volume is less than 0.5 mL, add working dialysis buffer to a
final volume of 0.5 mL (after the lower level of the meniscus has equilibrated).
0.75
0.5
0.2

11. Collect the dialyzed urine remaining in the tube and apply it in a 1.5 mL microtube.
of the dialysis and concentration device (SEBIA dialysis system), it is not recommended to store the sample in the dialysis system after centrifugation
but in a microtube stored refrigerated (between 2 and 8 C).
The instructions for the use of dialysis and concentration devices with swing bucket centrifuge and fixed angle centrifuge are different. The parameters
described above are indicated for the use of SEBIA dialysis systems with swing bucket centrifuge. Please see carefully the instruction sheets of each
dialysis and concentration device to use it with fixed angle centrifuge.
Samples to avoid
Do not re-use the urine sample applied in the reagent cup.
- 33 -
PROCEDURE
The MINICAP system is a multiparameter instrument to analyze urinary proteins on 2 parallel capillaries. The sequence of automated steps is as
follows :
Bar code reading of samples to analyze (for up to 26 tubes) and rotating sampler ;
Samples application into reagent cups from microtubes containing the dialyzed samples ;
Capillary washing ;

Sample preparation (dialysis and concentration) ;
Set up microtubes (without caps) containing the samples to analyze on holding hemolysing tubes in rotating sampler in positions No. 1 to 26 ; each
tube being identified with the specific sample identification bar code label corresponding to the sample to analyze.
Set up the Normal Control Serum in rotating sampler in position No. 28.
Set up the rotating sampler in the MINICAP instrument ;
Removal after analysis of rotating sampler and microtubes for potential analysis by immunotyping.
Remove and close the bins for used cups
PLEASE CAREFULLY READ THE MINICAP INSTRUCTION MANUAL.
1. Switch on MINICAP instrument and computer.

2. In order to start the instrument, position at least one new reagent cup on the automated loading system for cups of MINICAP (a message will
be displayed if a reagent cup is missing).
3. Set up the software, the instrument automatically starts.
4. The MINICAP instrument must be used with "URINE" analysis program and the MINICAP PROTEIN(E) 6 buffer. To select "URINE" analysis
program and place the MINICAP PROTEIN(E) 6 buffer vial in position "B1" in the instrument, please read carefully the MINICAP instruction
manual and follow the instructions displayed on the screen.
5. Position new reagent cups on the automated loading system for cups of MINICAP (a message will be displayed if the reagent cups are
missing).
6. Position a new bin for used cups in MINICAP at the location intended for this purpose.
7. Check the filling level of the reagent vials, add reagent if necessary and empty the waste container. In the window "Check reagent levels",
update the software by moving the cursor buttons.
8. The rotating sampler contains 28 positions for tubes. To analyse the urine samples prepared with the CAPILLARYS / MINICAP URINE kit :
Place up to 26 empty hemolysing tubes (used as holders) on the rotating sampler (positions No. 1 to 26) by starting into position No. 1,
and then, the microtubes containing the dialyzed urine samples. Cut the cap of each microtube before using it. The bar code of each tube
must be visible in the openings of the rotating sampler.
IMPORTANT : It is recommended to identify each hemolysing tube holding the microtube which contains samples to analyze, with the
specific sample identification bar code label corresponding to the sample. This identification allows to determine the necessary dilution
program to use when performing the MINICAP IMMUNOTYPING URINE technique. When bar codes are missing, use the software function
that allows to link the position of the tube on the rotating sampler with the dilution program selected for the MINICAP IMMUNOTYPING
URINE procedure.
Keep the cap of each microtube for further storage of samples, if necessary.
If the number of samples to analyze is odd, add a tube containing distilled or deionized water.
To normalize the capillaries with the Normal Control Serum (see the paragraph "Reagents required but not supplied"), apply 25 L
reconstituted Normal Control Serum in a hemolysing tube and 2 mL of working dialysis buffer ; then, homogenize and position this tube in
position No. 28 on the rotating sampler ("Control" position).
IMPORTANT : It is necessary to normalize the capillaries before urine samples analysis in the MINICAP instrument in order to analyze them
afterwards with the MINICAP IMMUNOTYPING URINE procedure. The results are automatically considered by the software for the data
analysis.
IMPORTANT : If a tube is missing in the position No. 1 (urine sample) and in the position No. 28 (Normal Control Serum), the analysis can
not start and a message will be displayed.
NOTE : When using the Normal Control Serum, it is necessary to use the specific bar code label.
9. Slide the rotating sampler into the MINICAP system.

10. Close the doors of the MINICAP system and start the analysis.
11. After the analysis, remove the rotating sampler and microtubes containing the analyzed urine samples and store them refrigerated (between
2 and 8 C) for further analysis with MINICAP IMMUNOTYPING URINE technique, if necessary.
12. If necessary, take off carefully the bin containing used reagent cups, close it tightly with the corresponding lid and discard it.
WARNING : Bins containing used reagent cups with biological samples have to be handled with care.
1.
2.
3.
4.
5.
6.
Bar codes are read on tubes with urine samples to analyze and on rotating sampler.
Samples are applied into reagent cups and the sample probe is rinsed after each sample.
NOTE : These automated steps are described for the two first analyzed sample tubes. The electrophoretic patterns appear after about 10 minutes
from the start of the analysis. For the following sample tubes, the two first steps (bar code reading and sample application) are performed during the
analysis of the 2 previous sample tubes.
- 34 -
II. RESULT ANALYSIS
NOTE : For each of the 3 sample preparation procedures, the extent of the pattern visualized on the y-axis on the screen takes into account the total
Abnormal fractions must be integrated using the "quantify peaks" tool of the software, then, the "Ratio urine", defined by the proportion of the abnormal
fraction area related to the Normal Control Serum proteins total area, is automatically calculated ; it will be displayed in the modify curves window.
This ratio will be used in the MINICAP IMMUNOTYPING URINE technique to determine the sample dilution program to use ("HYPOGAMMA",
"STANDARD" or "HYPERGAMMA").
At the end of each analysis sequence, the operator must initiate the "stand by" or "shut down" procedure of the MINICAP System in order to store
IMPORTANT : Position at least one new reagent cup on the automated loading system for cups of MINICAP (a message will be displayed if a reagent
cup is missing).
The MINICAP System has a reagent automatic control.

IMPORTANT : Please refer to the instructions for replacement of reagent containers respecting the color code for vials and connectors.
RESULTS
Interpretation
As an aid for patterns interpretation, it is recommended to overlay a pattern obtained after the normalization of capillaries with the Normal Control
Serum to obtain a reference pattern.
2.0 mg/dL.
An identification is recommended to characterize monoclonal or oligoclonal components :
- by immunotyping with SEBIA MINICAP IMMUNOTYPING URINE technique or,
- by immunofixation with SEBIA HYDRAGEL BENCE JONES, HYDRAGEL IF and HYDRAGEL URINE PROFIL(E) kits and specific antisera anti-free
kappa and lambda light chains.
Particular cases :
whole pattern.

Analyze only samples prepared with dialysis and concentration devices, indicated in the paragraph "EQUIPMENT AND ACCESSORIES REQUIRED",
such as SEBIA dialysis systems or equivalent device giving the same performances and approved for clinical assays.
- 35 -

sample again.
Troubleshooting
carefully followed.
PERFORMANCE DATA
Standard materials, sample preparation and procedures were used. All electrophoregrams were evaluated visually.
Concordance Study
Concordance study was performed on 46 different urine samples between MINICAP URINE procedure and a commercially available capillary
electrophoresis system for urine analysis : 1 normal urine sample and 45 different pathological urines have been run on both systems ; the total protein
concentrations were comprised between 0.005 and 1.196 g/dL.
This study demonstrated a 100 % agreement between the two techniques :
For the normal urine sample : complete agreement (concordance).
For the 45 pathological urine samples : complete agreement (concordance).
For all the analyzed pathological urine samples, the results obtained with the individual procedures were in agreement and the urinary protein fractions
were detected on all pathological samples with each system.
- 36 -
BIBLIOGRAPHIE / BIBLIOGRAPHY
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
Cameron JS. 1987. The nephrotic syndrome. Am J Kidney Dis, 10 : 157-171.

Chopin N. 1991. tude de la protinurie. Inf Tech Biol, 1 : 23-28.
Henskens Y et al. Detection and identification of monoclonal gammopathies by capillary electrophoresis. Clin Chem, 44, 1184-1190 (1998).
Jellum E et al. Diagnostic applications of chromatography and capillary electrophoresis. J Chromatogr B, 689, 155-164 (1997).
Joachim GR, Cameron JS, Chwartz M, Belker EL. 1964. Selectivity of protein excretion in patients with the nephrotic syndrome. J Clin Invest,
43, 2332-2346.
Landers JP. Clinical Capillary Electrophoresis. Clin Chem, 41, 495-509 (1995).
Le Bricon T. 2002. Identification et dosage des proteines urinaires au laboratoire danalyses. Ann Biol Clin, 60 : 525-540.
Le Carrer D. 1990. Protinuries : Mise au point sur leur exploration biologique en 1990. LEurobiologiste, 190 : 395-405.
Le Carrer D, Nicolas A, Ducasse L. 1992. Lanalyse des protinuries au laboratoire de biologie en 1992. Revue franaise des laboratoires, 225 :
41-47.
Le Carrer D, Chopin N. 1994. Profil protique urinaire : Proposition dun protocole dexploration biologique des protinuries. Revue franaise des
laboratoires, 269 : 29-37.
Philipon C. 1989. Protines urinaires : Intrt clinique et interprtation. Technique et Biologie, 6 : 239-249.
Oda RP et al. Capillary electrophoresis as a clinical tool for the analysis of protein in serum and other body fluids. Electrophoresis, 18, 1715-1723
(1997).
Waller KV, Ward KM, Mahan JD, Wismatt DK. 1989. Current concepts in proteinuria. Clin Chem, 35 : 755-765.
Wendling A. Procdures de diagnostic ou de dpistage : Justification et validit dun test de diagnostic ou de dpistage-sensibilit-spcificit.
Impact-Internat, 1986 ; Sept : 93-97.
Westermeier R., Electrophoresis in Practice. A Guide to Theory and Practice, VCH Publishers, New York, NY, 1993.
- 55 -
SCHMAS / FIGURES
Figure 1
Urine normale
Normal urine sample
Zones protiques
Protein zones
Albumin(e)
Alpha 1
Alpha 2
Beta
Gamma
Alpha 2
Beta
Gamma
Figure 2
Urine pathologique avec 5 zones protiques
Pathological urine sample with 5 protein zones
Zones protiques
Protein zones
Albumin(e)
Alpha 1
- 56 -
SCHMAS / FIGURES
Figure 3
Urine pathologique avec protine de Bence Jones
Pathological urine sample with Bence Jones protein
Zones protiques
Protein zones
Albumin(e)
Alpha 1
Alpha 2
Beta
Gamma
protine de Bence Jones

Bence Jones protein
Figure 4
Urine pathologique avec 2 protines de Bence Jones

Pathological urine sample with 2 Bence Jones proteins
Zones protiques
Protein zones
Albumin(e)
Alpha 1
Alpha 2
Beta
Gamma
2 protines de Bence Jones

2 Bence Jones proteins
- 57 -
SCHMAS / FIGURES
Figure 5
Profil de rfrence : Srum de Contrle Normal dilu au 1/40
Reference pattern : Normal Control Serum diluted 40 times
Zones protiques
Protein zones
Albumin(e)
Alpha 1 Alpha 2 Beta
- 58 -
Gamma

Capillarys / Minicap Urine

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Capillarys / Minicap Urine

Uploaded by

Copyright:

Available Formats

CAPILLARYS / MINICAP URINE

CAPILLARYS / MINICAP URINE - 2012/05

PRINCIPLE OF THE TEST

REAGENTS SUPPLIED IN THE CAPILLARYS / MINICAP URINE KIT

Dialysis buffer (stock solution)

FOR OPTIMAL RESULTS :

Storage, stability and signs of deterioration

SEBIA INSTRUCTIONS - English

CAPILLARYS / MINICAP URINE - 2012/05

CAPILLARYS URINE PROCEDURE

1. CAPILLARYS PROTEIN(E) 6 KIT (SEBIA, PN 2003)

Presentation, use, storage, stability and signs of deterioration

2. DISTILLED OR DEIONIZED WATER

Storage, stability and signs of deterioration

4. SODIUM HYPOCHLORITE SOLUTION (for sample probe cleaning)

Storage, stability and signs of deterioration

5. CAPILLARYS / MINICAP WASH SOLUTION

See instruction sheet for details.

Storage, stability and signs of deterioration

CAPILLARYS / MINICAP URINE - 2012/05

EQUIPMENT AND ACCESSORIES REQUIRED

CAPILLARYS System SEBIA : CAPILLARYS PN 1220 or CAPILLARYS 2 PN 1222.

SAMPLES FOR ANALYSIS

Sample collection and storage

7. Add 10 mL working dialysis buffer per dialysis system.

CAPILLARYS / MINICAP URINE - 2012/05

The manual steps include :

PLEASE CAREFULLY READ THE CAPILLARYS INSTRUCTION MANUAL.

I. PREPARATION OF ELECTROPHORETIC ANALYSIS

1. Switch on CAPILLARYS instrument and computer.

DILUTION - MIGRATION - DESCRIPTION OF THE AUTOMATED STEPS

CAPILLARYS / MINICAP URINE - 2012/05

II. RESULT ANALYSIS

III. END OF ANALYSIS SEQUENCE

IV. FILLING OF REAGENT CONTAINERS

The CAPILLARYS System has a reagent automatic control.

CAPILLARYS / MINICAP URINE - 2012/05

Interference and Limitations

See SAMPLES FOR ANALYSIS.

CAPILLARYS / MINICAP URINE - 2012/05

CAPILLARYS 1 & 2 release 6.x

1. CAPILLARYS PROTEIN(E) 6 KIT (SEBIA, PN 2003)

Presentation, use, storage, stability and signs of deterioration

2. NORMAL CONTROL SERUM

Storage, stability and signs of deterioration

3. DISTILLED OR DEIONIZED WATER

Storage, stability and signs of deterioration

5. SODIUM HYPOCHLORITE SOLUTION (for sample probe cleaning)

CAPILLARYS / MINICAP URINE - 2012/05

Storage, stability and signs of deterioration

6. CAPILLARYS / MINICAP WASH SOLUTION

See instruction sheet for details.

Storage, stability and signs of deterioration

EQUIPMENT AND ACCESSORIES REQUIRED

CAPILLARYS System SEBIA : CAPILLARYS PN 1220 or CAPILLARYS 2 PN 1222.

SAMPLES FOR ANALYSIS

Sample collection and storage

CAPILLARYS / MINICAP URINE - 2012/05

Sample preparation (with SEBIA dialysis systems)

Quantify total protein concentration in the urine sample to be analyzed.

7. For dialysis, add 20 mL working dialysis buffer per dialysis system.

10. Homogenize the sample in the tube.

CAPILLARYS / MINICAP URINE - 2012/05