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L-门冬酰胺酶(EC3.5.1.1)是一种专一性水解胞外L-门冬酰胺的蛋白酶,该酶主要用于治疗小儿急性淋巴细胞白血病。目前对该产品的鉴别方法之一是收录在2015年版《中国药典》中的反相高效液相色谱法(RP-HPLC)。研究门冬酰胺酶样品在反相分析方法的流动相中,样品结构和活性变化情况,作者收集了门冬酰胺酶样品在RP-HPLC色谱法中出峰阶段的组分,并分析其生物活性和蛋白一级结构。分析检测结果表明,门冬酰胺酶样品在反相分析方法的流动相中完全失去生物活性,但其蛋白一级结构没有改变。
L-asparaginase (EC3.5.1.1) is a proteolytic enzyme that specifically hydrolyzes extracellular L-asparagine, which is mainly used to treat childhood acute lymphoblastic leukemia. Currently, one of the methods for identifying this product is RP-HPLC, which is included in the 2015 edition of the Chinese Pharmacopoeia. To study the changes of the structure and activity of asparaginase samples in the mobile phase of RP-HPLC method, we collected the components of asparaginase sample in the peak phase of RP-HPLC and analyzed its bioactivity And protein primary structure. Analysis of the test results showed that asparaginase samples completely lost their biological activity in the mobile phase of the reverse phase analysis method, but the primary structure of the protein did not change.