Professional Documents
Culture Documents
Immunohaematology I
Lesson 9: Antihuman Globulin
Reaction
Immunoglobulin
Reagents
1. Monoclonal (single clone of cells-specificity)
2. Polyclonal (human source: mixture of cellscontains multiple antibodies)
Assist in identifying antigen present on
patients RBC that may cause and reaction in
vivo
Polyspecific AHG
Abs to human IgG and Abs to human C3d (antiC3d to detect Ab bound with complements
and those rare Abs)
Advantage is that polyspecific AHG may detect
complement-dependent Abs on RBCs
Monospecific AHG
Abs to human IgG or human C3d only
May miss an important antibody
Zeta Potential
An expression of the difference in electrostatic
potential at the surface of the red cell and the
ionic cloud of positive cations that are
attracted to the negative charges on the
surface
The net negative charge surrounding red cells
is part of the force that repels red cells from
each other
Enhancement Media
Low Ionic Strength Solution (LISS)
Decrease the ionic strength of a reaction medium
Thus, reduce the zeta potential
Leads to an increase in the attraction between
positively charged Ab molecules and negatively
charged red cells
Increase the rate of antibody uptake incubation
time of 5-15 mins; instead of 30-60 mins
Albumin
Enhances antibody detection test by reducing
the zeta potential
Allow Ab-sensitized cells to become closer
together
Do not promote the antibody uptake stage of
agglutination
Enzymes
Can enhance or suppress the reaction of
certain blood group antigens and antibodies
There are certain enzymes can be used:
Ficin (Fish)
Trypsin (pig stomach)
Papain (papaya)
Bromelin (pineapple)
Action of Enzyme
The treatment of red cells with enzyme
results in the release of sialic acid from the
membrane of RBC
Subsequent decrease in the negative charge
of the cells which lead to reduction in the
zeta potential
Reagent
Bovine serum albumin
(BSA)
Uses
Rh antibodies enhanced
at 37C