Nims R W, Shoemaker A P, Bauernschub M A, Rec L J, Harbell J W
MA BioServices, Inc., Rockville, Maryland 20850, USA.
In Vitro Cell Dev Biol Anim. 1998 Jan;34(1):35-9. doi: 10.1007/s11626-998-0050-9.
The analysis of the gel electrophoresis banding patterns and relative migration distances for the individual isoforms of intracellular enzymes, such as lactate dehydrogenase, purine nucleoside phosphorylase, glucose-6-phosphate dehydrogenase, and malate dehydrogenase, is used routinely in the biopharmaceutical industry for confirmation of cell line species of origin. In the present study, the sensitivity of the technique (AuthentiKit, Innovative Chemistry, Marshfield, MA) for determining interspecies cell line cross-contamination was examined. Extracts were prepared from a CHO-K1 line (AA8, Chinese hamster), MRC-5 (human) cells, and L929 (mouse) cells and from several proportional mixtures of the various binary combinations of cells. The isoenzymes were analyzed according to standard procedures for the technique. Contamination of MRC-5 cells with CHO-K1 or with L929 cells was clearly detectable with each enzyme analyzed. Similarly, the contamination of L929 or CHO-K1 cells with MRC-5 cells was readily apparent with each enzyme. On the other hand, contamination of CHO-K1 cells with L929 cells was only detected with lactate dehydrogenase analysis, and contamination of L929 cells with CHO-K1 cells was not detected with any of the four enzymes examined. For the latter case, the analysis of an additional enzyme (peptidase B) was required. The results indicate that interspecies cross-contamination should be detectable with isoenzyme analysis if the contaminating cells represent at least 10% of the total cell population.
对细胞内酶的各个同工型，如乳酸脱氢酶、嘌呤核苷磷酸化酶、葡萄糖-6-磷酸脱氢酶和苹果酸脱氢酶的凝胶电泳条带模式和相对迁移距离进行分析，在生物制药行业中常用于确认细胞系的起源物种。在本研究中，检测了该技术（AuthentiKit，Innovative Chemistry，马什菲尔德，马萨诸塞州）用于确定种间细胞系交叉污染的灵敏度。从CHO-K1细胞系（AA8，中国仓鼠）、MRC-5（人）细胞和L929（小鼠）细胞以及几种细胞二元组合的比例混合物中制备提取物。按照该技术的标准程序分析同工酶。分析的每种酶都能清晰检测到MRC-5细胞被CHO-K1或L929细胞污染。同样，每种酶都能很容易地看出L929或CHO-K1细胞被MRC-5细胞污染。另一方面，仅通过乳酸脱氢酶分析检测到CHO-K1细胞被L929细胞污染，而在所检测的四种酶中均未检测到L929细胞被CHO-K1细胞污染。对于后一种情况，需要分析另一种酶（肽酶B）。结果表明，如果污染细胞占总细胞群体的至少10%，则通过同工酶分析应可检测到种间交叉污染。
