重组胰高血糖素样肽-1受体激动剂(rExendin-4)原液质量研究

Study on the quality of bulk solution of recombinant pancreatic glucagon-like peptide-1 receptor agonist( rExendin-4)

目的:对重组胰高血糖素样肽-1受体激动剂(r Exendin-4)原液进行质量研究。方法:采用紫外光谱、肽图、飞行时间质谱及N端氨基酸序列分析对r Exendin-4原液进行结构确证;采用凯氏定氮法、BCA法及氨基酸分析法测定r Exendin-4原液的蛋白含量,建立高效液相色谱法测定原液纯度,通过纯度校正计算原液中r Exendin-4含量。结果:确证了r Exendin-4原液的结构,测得原液含量为2.21 mg·m L-1。结论:本研究所用的r Exendin-4原液结构正确、测定含量结果准确可靠,可作为HPLC外标法含量测定用对照品,用于r Exendin-4同质对照品的赋值,也可作为理化对照品用于r Exendin-4工艺中间体的评价和原液放行检验。

Objective： To study the quality of rExendin-4 bulk solution. Methods： UV, peptide mapping, TOF MS and N terminal amino acid sequencing were used to confirm the structure of rExendin-4 bulk solution Kieldahl method, BCA method and amino acid analysis method were applied to determine the protein content. The purity of rExendin-4 bulk solution was determined by HPLC. The content of rExendin-4 was calculated by purity correction. Results： The structure of the material was confirmed and the content was determined as 2.21 mg·mL-1 Conclusion： The established rExendin-4 bulk solution can be used as the reference substance for the con- tent assay of rExendin-4 by HPLC, value assignment of the and release test of rExendin-4 bulk solution homogeneous standard, evaluation of drug intermediate,