Isoenzyme Analysis
Before evaluating the biosafety of cell-derived products, manufacturers need to check and to guarantee the origin of the cells used, as described in the FDA and EMEA guidelines.
Three cell banks are generally examined: the master cell bank (MCB), the working cell bank (WCB) and the end-of-production cell bank (EPCB) or cells at the age limit of in vitro culture.
The tests carried out depend on the origin of the cells in question. Isoenzyme analysis is generally requested but if the cell line is derived from a heterohybridoma (e.g. human/mouse), DNA fingerprinting is sometimes required.
Protocol 0302: Isoenzyme Analysis |
Product to be tested: cell banks |
This protocol is intended to verify cell line identity and to detect cell line contamination by comparing the migration distances of four different isoenzymes (chosen from the following: glucose-6-phosphate dehydrogenase (G6PD), nucleotide phosphorilase (NP), malate dehydrogenase (MD), lactate dehydrogenase (LD), aspartate aminotransferase (AST), mannose phosphate isomerase (MPI), peptidase B (PEP B)) in cell extracts after electrophoresis and activity staining. The analysis provides a permanent record of the cell line isoenzyme profile, which is unique to each animal species. |
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