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  

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

作　　者：张竞 周志艳 李丹丹 安丽康 王岩勃 邹卫[4] ZHANG Jing;ZHOU Zhiyan;LI Dandan;AN Likang;WANG Yanbo;ZOU Wei(Hebei Jizhaolian Science and Technology Development Co.Ltd.,Shijiazhuang050051,Hebei Province,China;不详)

机构地区：[1]河北冀兆联科技发展有限责任公司,河北石家庄050051 [2]河北省药品医疗器械检验研究院,河北石家庄050200 [3]石家庄沃泰生物科技有限公司,河北石家庄050018 [4]河北科技大学,河北石家庄050018

出　　处：《中国生物制品学杂志》2023年第5期599-603,613,共6页Chinese Journal of Biologicals

基　　金：中央引导地方科技发展资金(226Z2404G)。

摘　　要：目的建立重组人脑利钠肽等电点(isoelectric point,pI)分析的全柱成像毛细管等电聚焦电泳(capillary isoelectric focusing-whole column imaging detection,CIEF-WCID)法,并进行验证。方法采用CIEF-WCID法对两性电解质、占位剂、蛋白浓度、聚焦时间等进行优化后,获得检测重组人脑利钠肽的最佳检测条件。对建立的方法进行重复性、样品间精密度和耐用性验证,并对连续生产的3批重组人脑利钠肽进行pI分析。结果选择HR AESlyte 8-10.5为两性电解质,25 mmol/L氢氧化钠为占位剂,以87.5μg/mL为样品检测终浓度,聚焦时间为8 min。同一样品连续进样6次,检测pI相对标准偏差(RSD)为0.1%;6份样品检测pI RSD为0.1%;在样品不同终浓度下,主峰pI RSD为0.1%,不同放置时间下,样品pI RSD为0.1%,但pI标记物不可随意更改。连续3批重组人脑利钠肽样品pI均可检出。结论建立的重组人脑利钠肽pI分析的CIEF-WCID法,重复性好、精密度高,可用于重组人脑利钠肽后续的质量控制。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 87.5μg/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.

关 键 词：重组人脑利钠肽 等电点 全柱成像毛细管等电聚焦电泳 优化 验证 

分 类 号：R917[医药卫生—药物分析学]