猪内脂素重组蛋白溶液中内毒素的去除  

Removal of Endotoxin from Swine Visfatin Recombinant Lipoprotein Solution

作　　者：许粉亮 李慧珍 杨文杰[1] 董翎 牛晓玉 宋卉[1] XU Fenliang;LI Huizhen;YANG Wenjie;DONG Ling;NIU Xiaoyu;SONG Hui(Huazhong Agricultural University,Wuhan 430070,China)

机构地区：[1]华中农业大学,武汉430070

出　　处：《中国畜牧兽医》2020年第12期3890-3896,共7页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金面上项目(31772687);中央高校基本科研业务费专项基金(2662015PY063);国家自然科学基金青年基金项目(31101776)。

摘　　要：本试验旨在探索4℃条件下蛋白的上样体积、流速和蛋白浓度对猪内脂素重组蛋白溶液中内毒素的去除效果和蛋白损失量的影响。该研究采用商品化、修饰过的多黏菌素B为介质,特异性地去除蛋白溶液中的内毒素。在4℃条件下,选用不同的上样体积(6、8、10、12 mL)、流速(0.125~0.75 mL/min)和蛋白浓度(0.9、1.2、1.3、1.7 mg/mL)进行内毒素的去除,使用内毒素检测试剂盒定量测定内毒素含量,使用BCA蛋白浓度测定试剂盒进行蛋白浓度的测定。试验结果表明:在4℃条件下,蛋白上样体积12 mL、流速0.125~0.25 mL/min时,内毒素的去除效果最佳,去除率高达99.5%,内毒素的残余量(1 EU/mL)在生物安全范围以内,并且蛋白的回收率高达90~95%。试验结果证明成功去除猪内脂素重组蛋白溶液中的内毒素,为该蛋白的相关功能研究和临床应用提供参考。The purpose of this experiment was to explore the influence of the sample loading volume,flow rate and protein concentration of protein on the removal efficiency of endotoxin and protein loss in porcine visfatin recombinant lipoprotein solution at 4℃.In this study,commercial and modified polymyxin B was used as the medium to specifically remove endotoxin from the protein solution.At 4℃,different loading volume(6,8,10,12 mL),flow rate(0.125-0.75 mL/min)and protein concentration(0.9,1.2,1.3,1.7 mg/mL)were used for endotoxin removal,the content of endotoxin was determined by a quantitative method using endotoxin reagents,and the BCA protein concentration assay kit was used to determine the protein concentration.The results showed that at 4℃endotoxin removal was the best when protein loading volume was 12 mL and flow rate was 0.125-0.25 mL/min,the removal rate was 99.5%,the residual amount of endotoxin(1 EU/mL)was within the biosafety range,and the protein recovery rate was as high as 90%to 95%.The experimental results showed that the endotoxin was successfully removed from the solution of porcine lipin recombinant protein,which provided references for the related functional research and clinical application of the protein.

关 键 词：猪内脂素重组蛋白溶液 内毒素 去除条件 去除率 回收率 

分 类 号：Q593.1[生物学—生物化学]