Seed Lot System

Definition Importance Necessity

By

M.Lotfi, D.V.M, Ph.D.

1389/9/23

Definition
In order to prevent the unwanted drift of properties which might ensue from repeated subcultures or multiple generations, the production of biological medicinal products obtained by microbial culture, cell culture or propagation in embryos and animals should be based on a system of master and working seed lots and/or cell banks.

Production of Vaccine Master seed

GMP

Working seed Inoculum

Media/Cell culture
Excipients Validation - process - method Stability studies

Single harvest

Pool/Concentrated material Purified/Bulk material Final lot Final bulk

Importance of Seed
Safe Seed

Good product

Potent

Seed

Cost Benefit

Seed

Importance of seed lot system

1. 2. 3. 4. 5. 6.

To have Safe, Pure and effective products To have valid QC test To compliance with cGMP Permanent production To have Good R&Ds Security

Speciality of Seed Lot System

1.

2.

3.

The fabrication of biological product is based on a system of master and working seed lots and cell banks. Seed lots and cell banks are established in a suitably controlled environment to protect the seed lot or the cell bank and the personnel. Seed lots and cell banks are characterized according to ICH Q5D and maintained in such a way as to minimize the risks of contamination or alteration.

Speciality of Seed Lot System

Continued
4.

The stability of seed lots and cell banks is documented, including the efficiency of recovery. Storage containers are appropriate to their function ( Storage containers should be hermetically sealed), clearly labeled (Barcode) , and kept at a suitable temperature. Storage temperatures are properly monitored. Deviations from set limits and any corrective actions are evaluated and documented.

Speciality of Seed Lot System

Continued
5.

6. 7.

8.

Only authorized personnel under the supervision of a responsible person, may handle seed lots and cell banks. An inventory is meticulously kept. (FIMS) A system is in place to ensure security and retrievability of seed lot or cell bank vials, without confusion or cross-contamination. All vials of master or working cell banks that removed from storage, are not returned.

Strategy in Seed Lot System

1.

2.

3.

The cell bank system consists of two tiers: a master cell bank (Master cell bank established from single clone) (MCB); and a working cell bank (WCB). should stored the MCB and WCB in two or more separate locations within the facility. To reduce the possibility of phenotypic variations, genetic drift, and contamination as much as possible, the number of passages should be minimized (five passages ) .
Master chart.doc Master seed felow chart.doc

4.

Extensive characterization of the MCB Because all WCBs would be derived from the well-characterized MCB.

ND vaccine

OIE

EP
Purity

CFR
1.Bacteria and fungi 1.Bacteria 2.Mycoplasma 2.Mycoplasma 3.Avian origin vaccine 3.Avian Salmonella contamination Lymphoid leukosis HemagglutinatingExteraneous viruses 4.Extraneous viruses 4.Extraneous by the chicken embryo inoculation test by the chicken inoculation test

Seed

Pathogenicity

Index of neurovirulence Immunogenicity

Safety Identity
1.Fluorescent antibody test 1.Fluorescent 2.Serum neutralization test 2.Serum Cell culture requirements

Stability Immunogenicity

In process control Batch control

Potency Viability Sterility Sterility Safety Potency

( by titration of infectivity)

Duration of immunity Stability Safety Potency Identity

Final product

Safety Potency

By HA Or cell cultures Or fertilised hen eggs Extraneous viruses 1.using cell cultures 1.using 2.using fertilised eggs 2.using 3.using chicks 3.using 4.Avian leucosis viruses 4.Avian

Virus titer Sterility Mycoplasma

Purity 1.Bacteria and fungi 1.Bacteria 2.Mycoplasma 2.Mycoplasma 3.Avian origin vaccine 3.Avian Salmonella contamination Lymphoid leukosis Hemagglutinating Extraneous viruses 4.Extraneous viruses 4.Extraneous By the chicken inoculation test By the chicken embryo inoculation test Safety Virus titer

Seed Lot System References

1.

2.

3.

ICH Q5D: Derivation and Characterization of Cell Substrates Used for Production of Biotechnology/Biological Products. ICH Q5A: Viral Safety Evaluation of Biotechnology Products Derived from Cell Lines of Human or Animal Origin. ICH Q5B: Quality of Biotechnology Products: Analysis of the Expression Construct in Cells Used for Production of R-DNA Derived Proteins. 21 CFR 610.18

Necessity of Seed Lot System

1. 2. 3.

Manufacturers should describe the type of banking system used. size of the cell bank, container, preparation, storage Manufacturers should describe their cell banking procedures. Manufacturers should describe the procedures used to avoid microbial contamination and crosscontamination by other cell types present in the laboratory. documentation system, labelling system, storage and recovery
from storage (FIMS).

General Principles of Seed Characterization

Historical Record Identity Test Purity Test Safety Test Efficacy Test Stability Test

Kinds of Seed Banks

Cell Bank Viral Bank Microbial bank Protozoal Bank

Cell Bank characterization

Identity Test Purity Test Safety Test Stability Test Tumorigenicity Test

Microbial Bank characterization

Identity Test morphological, cultural, biochemical, genetic and
serological

Purity Test Safety Test Stability Test

Viral Bank characterization

Identity Test Purity Test Safety Test Stability Test

Facility For characterization of Seeds

Building Cell culture lab Microbiological lab Immunological lab Molecular lab Animal facility Virological lab