PRESENTER: K. RAJENDRA09-02-2013

MAINTENANCE OF CGMP

CELL BANKS

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

DESIGN, DEVELOP & DELIVER

Global CMC Support

Consultancy Regulatory

or Training

In-House Strategic Issues

Clinical logistics

Process design Cell Line Development

& Development Process Development

Analytical Development

Cell Banking

Cell Bank Storage

cGMP Manufacture Mammalian Cell Culture

Microbial Fermentation

Viral Production

DESIGN

DEVELO

P

DELIVER

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.

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.

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.

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 without loss of labelling information on the container.

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.

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.

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 containers1)

Tests for presence of Mycoplasma*.

In Vitro assay In Vivo assay

Sterility testing.

Viability testing.

• Tests described in either the Ph. Eur, JP or U.S.P

• 1- Q5D,/21CFR610.12.

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

DOCUMENTATION OF CELL BANKS & VALIDATION

New Culture

Master Cell Bank

Working Cell Bank

Cell Stocks at Extended Passage (e.g. every 10 to 20 passage)

Cell Line Data File

Accession No.

Cell Bank Ref. No

QC & Validation Ref.

Passage records

Validation data

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.

Access of cell bank should be restricted.

DOCUMENTATION OF STORED MATERIAL

Traceability through Reference Numbers on Laboratory Records.

Reference No

Accession Day Book

Cell Bank Ampoules

Cell Banking Records

Q.C. Records

Media Batch Preparation Record

Sterilization Record

Accession No. + + +Cell Bank No. + + +Q.C. Ref. No. + +Media Batch Ref. No. + + +

Sterilization Reference + +

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

“THANK YOU”

K. Rajendra

“GOOD SEEDS GIVES YOU SWEET FRUITS”