重组人抗血栓蛋白质控方法和质量标准研究

Methods and requirements for quality control of recombinant human antithrombus protein

目的:建立重组人抗血栓蛋白(recombinant human antithrombus protein,rHAP)的质控方法和质量标准。方法:采用活化部分凝血活酶时间法(APTT法)测定生物学活性;还原型SDS-PAGE测定相对分子质量;非还原SDS-PAGE和反相高效液相色谱测定纯度;胰蛋白酶酶切后用反相高效液相色谱法分析肽图;其余检测项目按2010年版《中华人民共和国药典》三部规定进行。结果:用建立的方法对原液和成品进行了检定,各项指标均符合《人用重组DNA制品质量控制技术指导原则》和2010年版《中华人民共和国药典》三部的要求。结论:建立的质控方法和质量标准能够保证产品安全、有效、质量可控,可用于注射用重组人抗血栓蛋白产品的常规检定。

Objective： To establish methods and requirements for quality control of recombinant human antithrombus protein （rHAP）. Methods： The bioactivity of rHAP was determined by APTT assay. Molecular weight was detected by reduced SDS-PAGE. Purity was analyzed by SDS-PAGE and RP-HPLC. Individual peptide mapping were detected by tryptic digestion. Other tests were performed according to Chinese Pharmacopeia. Results： The requirements and methods for quality control of rHAP had been established and used for determining both rHAP bulk and final product. The results of all tests fit Chinese Pharmacopeia and Guidelines for Quality Control of Recombinant DNA Products. Conclusion ： The methods and requirements for quality control of rHAP can assure safety and efficiency of the products, and can be used for routine quality control of rHAP.