This document outlines guidelines for biologics testing in Europe put forth by the EMEA and ICH. It discusses characterization that should be done at various stages of production including the master cell bank, working cell bank, end of production bank, bulk harvest, and final product. Testing includes sterility, mycoplasma detection, adventitious virus testing both in vitro and in vivo, identity testing, characterization of product attributes, and other quality control tests to ensure safety and efficacy.

1. Biologics testing in Europe
EMEA ICH Guidelines
CPMP/ICH/295/95/ICH Topic Q5A
CPMP/ICH/294/95/ICH Topic Q5D
CHMP/BWP/157653/07
EMEA/CHMP/BWP/398498/05
Characterisation of MCB
Characterisation of Final Product Testing
3AB2A CPMP/ICH/139/95 ICH Topic Q5B Bulk Harvests CPMP/ICH/365/96 ICH Topic Q6B
CPMP/BWP/1143/00
Master Cell Bank Working Cell Bank End of Production Bank
• Morphology - light microscope • Morphology – light microscope • Morphology – light microscopic • Sterility conforms to EP 2.6.1 or • Sterility conforming to EP 2.6.1
observations observations observations Bioburden conforming to EP 2.6.12 • pH conforms to EP 2.2.3
• Identity – DNA Fingerprinting • Identity – DNA Fingerprinting • Identity – DNA Fingerprinting • Mycoplasma complies with EP 2.6.7 or • Visible and sub-visible particles
EP 2.6.21 conforming to EP 2.9.19
• Sterility conforming to EP 2.6.1 • Sterility conforms to EP 2.6.1 • Sterility conforms to EP 2.6.1
• in vitro adventitious virus testing – • Extractable volume conforms to EP
• Mycoplasma test which complies with • Mycoplasma test which complies with • Mycoplasma test (EP 2.6.7) MRC-5, Vero and cell line of the same 2.9.17
EP 2.6.7 EP 2.6.7
• in vitro adventitious virus test using species and tissue type as the production
• in vitro adventitious virus test using Simian Simian (Vero), Human (MRC-5)and cell line (14 day assay in the case of • Bacterial endotoxins conforms to EP
(Vero), Human (MRC-5) and cells of cells of the same species and tissue non-primate cells, 28 day assay for 2.6.14
the same species and tissue type as the type as the production cells primate cells) • Total proteins conforming to EP 2.5.33
production cells, (14 day assay in the case
• in vivo test in adult and suckling mice • Tests for Retroviruses including PERT • Residual Host cell DNA
of non-primate cells, 28 day assay for
and embryonated chickens eggs (following EP 2.6.21), Transmission • Residual Host cell Protein
primate cells)
• Tests for Retroviruses including PERT Electron Microscopy, Infectivity assays
• in vivo adventitious virus test in adult and • Characterisation of product
(following EP 2.6.21), Transmission • Tests for species specific testing i.e.
suckling mice and embryonate chickens • Molecular distribution – SEC HPLC
Electron Microscopy, Intfectivity assays Mouse minute virus or Cache Valley virus
eggs (may also include Guinea Pigs conforming to EP 2.2.10
i.e. PG4 S+L- or cocultvation • Once only – in vivo adventitious
where appropriate)
virus testing in adult and suckling • Purity – Gel Electrophoresis conforming
• Species specific tests inc. Bovine and to EP 2.2.30.
mice and embryonated chickens eggs
Porcine viruses (CPMP/BWP/1793/02)
(may also include Guinea Pigs where • Product Potency - in vivo or in vitro
• Tests for Retroviruses including PERT appropriate) assay following EP section 5.3
(following EP 2.6.21), Transmission
Electron Microscopy, Infectivity assays i.e. • (Biological Activity per mass unit)
PG4 S+L- or cocultvation • (CHMP/BWP/271475/06)
• Appearance conforming to EP 2.2.2
• Osmolality conforms to EP 2.2.35
• Product specific Testing requirement
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