This document summarizes the key aspects of maintaining current good manufacturing practices (cGMP) for cell banks. It discusses the design, development, and delivery of cell banks. It defines cGMP and describes the cell banking system including master and working cell banks. It outlines the characterization, documentation, storage, and validation requirements for cell banks including testing for identity, purity, and stability. The goals are to ensure consistent production of safe, pure, and potent biotherapeutic products through well-characterized and validated cell banks.

1. MAINTENANCE OF CGMP
CELL BANKS
PRESENTER: K. RAJENDRA
09-02-2013

2. TABLE OF CONTENT
• Design, Develop & Deliver
• cGMP Definition
• Cell Banking
• Characterization of Cell Banks
• Tests for Identity
• Tests for Purity
• Tests for stability
• Documentation of Cell Banks and Validation
• Cell Bank Storage
• Documentation of stored material

3. DESIGN, DEVELOP & DELIVER
Global CMC Support
Consultancy Regulatory IGN
or Training DES
In-House Strategic Issues
Clinical logistics
LO P
EVE
Process design Cell Line Development
& Development Process Development D
Analytical Development
VER
Cell Banking
Cell Bank Storage
DELI
cGMP Manufacture Mammalian Cell Culture
Microbial Fermentation
Viral Production

4. CURRENT GOOD MANUFACTURING
PRACTICES (CGMP)
DEFINITION
“A set of current, scientifically sound methods, practices
or principles that are implemented and documented
during product development and production to ensure
consistent manufacturer of safe, pure and potent
products”.
FDA - Guidance for Industry: cGMP for Phase 1 Investigational Drugs.

5. CELL BANKING
(ICH Q5D / EMEA)
Cell bank – A cell bank is a collection of appropriate
containers, whose contents are of uniform composition,
stored under defined conditions. Each containers
represents an aliquot of a single pool of cells.
MCB (Master Cell Bank) – An aliquot of a single pool of
cells prepared from the selected clone under defined
conditions. The MCB is used to derive working cell banks.
WCB (Working Cell Bank) – The Working Cell Bank is
prepared from aliquots of a homogenous suspension of
cells obtained from culturing the MCB under defined
culture conditions.

6. CELL BANKING SYSTEM
(ICH Q5D / EMEA)
The cell bank system consists of two tiers:
1.Master Cell Bank (MCB – Single tiered).
2.Working Cell Bank (WCB – Two tiered) or
Manufacturer’s Working Cell Bank (MWCB).
A newly prepared WCB should be appropriately
qualified by characterization and testing.

7. CELL BANKING PROCEDURES
(ICH Q5D / EMEA)
Manufacturers may prepare their own cell banks or
may obtain from external sources.
Manufacturers should describe the type of banking
system used along with size of cell bank, container and
closure system, cryopreservation and storage methods.
Manufacturers should describe the procedures used to
avoid microbial contamination and cross-
contamination by other cell types present in the
laboratory.
Labelling system which can withstand the process of
preservation, storage, and recovery from storage

8. CHARACTERIZATION OF CELL BANKS
(ICH Q5D / EMEA)
 Testing objectives of cell line
--Confirm identity
--Confirm purity
--Confirm Suitability or Stability
 Quality assurance established from master bank to
end-of-production/post production cells (EPC/PPC).
 The manufacturer may choose to characterise the
WCB instead of the MCB, if justified.

9. TESTS FOR IDENTITY
(ICH Q5D / EMEA)
 Phenotypic or Genotypic
 Generally performed on MCB
limited to WCB.
 For human or animal cells
 Morphological, Isoenzyme
analysis
 Banding cytogenetics or species
– specific antisera.
 For microbial cells
 Phage typing.

10. TESTS FOR PURITY
(ICH Q5D / EMEA)
 Free from adventitious microbial
agents and cellular contaminants.
 Generally performed on MCB &
WCB.
 Tests for presence of Bacteria &
fungi*.
(1% of total No. but not less that two containers 1)
 Tests for presence of Mycoplasma*.
 In Vitro assay
 In Vivo assay
 Sterility testing.
 Viability testing.

11. TESTS FOR STABILITY
(ICH Q5D / EMEA)
 Two concerns for cell bank stability tests.
 Consistent production of the intended product.
 Retention of production capacity during storage.
 Generally performed on MCB & WCB.
 Tests for Copy number determination
 DNA & RNA Sequencing
 Restriction map analysis
 Retention of selectable markers
 Retention of recombinant construct

12. DOCUMENTATION OF CELL BANKS &
VALIDATION
Accession No. New Culture
Master Cell
Bank
Passage records
Cell Bank Ref. No
Validation data
Working Cell
Bank
QC & Validation Ref.
Cell Stocks at Extended Passage (e.g.
every 10 to 20 passage)
Cell Line Data File

13. CELL BANK STORAGE
The cell bank storage should be:
Long term storage.
(e.g., liquid nitrogen, ultra-low temperature
freezer)
(or)
Vapor phase
(as compared to the liquid phase).
Cell stability under the freezing and storage conditions
should be validated using cell recovery or viability data.
Storage of MCB & WCB should be in two or more
locations.


14. DOCUMENTATION OF STORED
MATERIAL
Traceability through Reference Numbers on Laboratory
Records.
Reference No Accession Cell Bank Cell Q.C. Media Batch Sterilization
Day Book Ampoules Banking Records Preparation Record
Records Record
Accession No. + + +
Cell Bank No. + + +
Q.C. Ref. No. + +
Media Batch
Ref. No. + + +
Sterilization
Reference + +

15. REFERENCES
 ICH- Q5D, Derivation and characterisation of cell substrates used
for production of biotechnological / biological products, July
1997.
 FDA Guidance for Industry - Characterization and Qualification of
Cell Substrates and Other Biological Materials Used in the
Production of Viral Vaccines for Infectious Disease Indications,
Feb’ 2010.
 Glyn Stacey - Fundamental Issues for Cell Line Banks in
Biotechnology and Regulatory Affairs Cell Biology, NIBSC, South
Mimms, UK 2004.
 EMEA 2006: CPMH/ICH/294/95, Note for guidance on quality of
biotechnological products: derivation and characterisation of
cell substrates used for production of biotechnological/biological
products

16. “GOOD SEEDS GIVES YOU SWEET
FRUITS”
“THANK YOU”
K. Rajendra