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LABORATORY PROCEDURES FOR ANIMAL & HUMAN CELL LINES

REFERENCE NO: AHC/1998/3/3.1/1.2


TITLE: ISOENZYME ANALYSIS


Isoenzyme analysis (IEF) is based on the existence of enzymes with similar or identical specificity, but different molecular structures (isoenzymes). Using IEF of a defined set of enzymes a characteristic "mobility pattern" can be established for each specimen. If the isoenzymes from different specimens result in different electrophoretic mobilities, these samples can individually be distinguished from each other (1-3). We routinely use IEF for the speciation of cell lines as described in detail (4). In those cell lines described by the depositor as being derived from mammals, our emphasis is to exclude cross-contamination(s)/mix-ups between human and murine cell lines and to confirm the assumed species. Non-mammalian cell lines pass this identity control when their electrophoretic mobility pattern differs from that of human and murine cells. Furthermore, IEF can be used to exclude or confirm HELA-like contaminants in a suspicious human cell line (5).

Our IEF analysis utilizes up to 7 enzymes provided in the AuthentiKitTM-system from Innovative Chemistry, Marshfield, MA, USA (6).The following enzymes can be examined: AST (aspartate aminotransferase), G6PD (glucose-6-phosphate dehydrogenase), LDH (lactate dehydrogenase), MDH (malate dehydrogenase), MPI (mannose phosphatase isomerase), NP (nucleoside phosphorylase), PEP B (peptidase B).

References:

1. Montes de Oca F, Macy ML, Shannon JE: Isoenzyme characterization of animal cell cultures. Proc Soc Exp Biol Med 132: 462-469 (1969).

2. Macy ML: Identification of cell line species by isoenzyme analysis. TCA Manual 4: 833-836 (1978).

3. Halton DM, Peterson WD, Hukku B: Cell culture quality control by rapid isoenzymatic characterization. In Vitro 19: 16-24 (1983).

4. Steube KG, Grunicke D, Drexler HG: Isoenzyme analysis as a rapid method for the examination of the species identity of cell cultures. In Vitro Cell Dev Biol 31A: 115-119 (1995).

5. Gartler SM: Apparent HeLa cell contamination of human heteroploid cell lines. Nature 217: 750-751 (1968).

6. Innovative Chemistry: The AuthentiKit System. Handbook for Cell Authentication and Identification, 2nd Edition. Marshfield, MA (1988).


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