重组人干扰素α2b内毒素检查法的研究

Bacterial Endotoxin Test on Recombinant Human Interferon α2b

目的研究鲎试剂法与家兔法检测重组人干扰素α2b内毒素结果的相关性,并参照家兔热原试验的阈值量,来确定本制品质量标准中内毒素的限量标准。方法分别用不同浓度的内毒素溶液和含不同浓度内毒素的供试品溶液进行家兔热原质试验,确定供试品内毒素限值,并在此限值下进行内毒素检测,方法均参照2000年版《中国药典》和2000年版《中国生物制品规程》进行。结果在含不同浓度内毒素溶液的家兔热原质试验中,致热阈值小于5.0EUml。在含不同浓度内毒素供试品溶液的家兔热原质试验中,致热阈值小于1.25EUml。以此确定本品内毒素限值,检测该制品的内毒素含量,结果均符合规定。结论本品在内毒素含量小于3.5EU500万单位dose的范围内,根据具体的对照实验及制品的用途来规定细菌内毒素限值,可用鲎试剂法代替家兔法检测注射用重组人干扰素α2b冻干制剂的热原质。

Objective To study the relationship between the determination results of bacterial endotoxin by limulus test and rabbit test,and to determine the limit of bacterial endotoxin content in the product according to the threshold value defined in pyrogen test in rabbits.Methods Perform pyrogen tests in rabbits using the bacterial endotoxin solutions at different concentrations and the test sample solutions containing different concentrations of bacterial endotoxin to determine the limit of bacterial endotoxin content in the test sample.Determine the bacterial endotoxin content,within the limit,of recombinant human interferon α2b by the methods described in Chinese Pharmacopoeia (2000 edition) and Chinese Requirements for Biologics (2000 edition).Results The threshold value of bacterial endotoxin solution causing fever of rabbit in pyrogen test was less than 5.0 EU/ml,and that of test sample solution containing bacterial endotoxin was less than 1.25 EU/ml.The limit of bacterial endotoxin content in recombinant human interferon α2b was defined according to the threshold values.The bacterial endotoxin content in the product met the related requiremnts.Conclusion Recombinant human interferon α2b showed synergistic action with bacterial endotoxin in causing the fever of rabbits.The limit of bacterial endotoxin content should be determined according to the individual control test and the usage of the product.The product with a bacterial endotoxin content of less than 3.5 EU per 5×106 IU showed no interference on bacterial endotoxin test.Limulus test might be used for the pyrogen test on recombinant human interferon α2b instead of rabbit test.