Development and verification of capillary isoelectric focusing-wholecolumn imaging detection method for isoelectric point analysis of recombinant human brain natriuretic peptide

ZHANG Jing

白话标题:重组人脑利钠肽等电点分析全柱成像毛细管等电聚焦电泳法的建立及验证
作者: ZHANG Jing
出版类型:Journal Article
关键字: Recombinant human brain natriuretic peptide; Isoelectric point(pI); Capillary isoelectric focusing-whole column imaging detection(CIEF-WCID); Optimization; Verification
来源: Chinese Journal of Biologicals 2023;36(5):599-603+613
国家China
语言:Chinese
摘要: Objective To establish and verify a capillary isoelectric focusing-whole column imaging detection(CIEFWCID) method for the analysis of isoelectric point(pI) of recombinant human brain natriuretic peptide.Methods The ampholyte,space-occupying agent,protein concentration,focusing time were optimized by CIEF-WCID method,and the best condition for the detection of recombinant human brain natriuretic peptide was obtained.The repeatability,precision and durability of the developed method were verified,and three batches of recombinant human brain natriuretic peptide produced continuously were analyzed for pI.Results HR AESlyte 8-10.5 was selected as ampholyte,while 25 mmol/L sodium hydroxide as the space-occupying agent;The final concentration of the sample was 87.5 μg/mL and the focusing time was 8min.The relative standard deviation RSD of pI detection was 0.1% after six consecutive injections of the same sample;The RSD of pI detection of six samples was 0.1%;The pI RSD of the main peak was 0.1% at different final concentrations of the sample,and the pI RSD of the sample was 0.1% at different storage time,while the pI markers could not be changed arbitrarily.The pI was detected in three consecutive batches of recombinant human brain natriuretic peptide samples.Conclusion The developed CIEF-WCID method for pI analysis of recombinant human brain natriuretic peptide had good repeatability and precision and might be used for follow-up quality control of recombinant human brain natriuretic peptide.