利凡诺方法分离纯化犬血浆蛋白络合及解离条件探讨

实验动物科学 ›› 2012, Vol. 29 ›› Issue (06): 30-33.

利凡诺方法分离纯化犬血浆蛋白络合及解离条件探讨

( 1. 军事医学科学院实验动物中心，北京100071) ( 2. 解放军第302 医院动物实验中心，北京100039)

出版日期:2012-12-30 发布日期:2014-04-16
基金资助:
军事医学科学院实验动物中心青年基金( QN2012—005)

Investigation on Conditions of Complexation and Dissociation in Separating and Purifying Dog Plasma Proteins with Rivanol Method

Online:2012-12-30 Published:2014-04-16

摘要/Abstract

摘要： 摘要: 目的普通犬血浆和高免血清分别是白蛋白和特异性免疫球蛋白分离纯化的原材料，针对不同的血浆确立相应的络合条件和解离条件，以分离纯化白蛋白和丙种球蛋白。方法血浆按1 ∶ 1 体积加入利凡诺溶液，用2% Na2 CO3调pH 值，使犬血浆与利凡诺进行充分络合; 上清液两次络合后解离，获得丙种球蛋白; 沉淀解离获得白蛋白。分别对上清液、两次解离液进行蛋白浓度、纯度的测定。结果确定pH 8. 5，室温络合2 h，利凡诺浓度在 2. 3% 时为白蛋白的最佳络合条件; pH 6. 0，室温络合2 h，利凡诺浓度2. 3% 为丙种球蛋白最佳络合条件。0. 6% 氯化钠，0. 25% 辛酸钠，pH 1. 37 为白蛋白解离最佳条件; 0. 85% 氯化钠，1. 3% 甘氨酸，pH 6. 8 为丙种球蛋白的最佳解离条件。结论利用利凡诺方法，可以成功分离和纯化犬白蛋白和丙种球蛋白。

关键词: 利凡诺方法, 络合及解离, 犬血浆, 分离纯化

Abstract: Abstract: Objective To investigate conditions of complexation and dissociation in separating and purifying albumin and specific immunoglobulin from ordinary dog plasma and hyperimmune serum with rivanol method． Method The plasma and rivanol solution were mixed in the volume ratio of one to one and pH value of the mixture was adjusted with 2% Na2CO3 to make the dog plasma and the rivanol full complexation． The gamma globulin was obtained by adding rivanol solution to the supernatant for secondary complexation and dissociating with dissociation dissociation， and the albumin was obtained from the precipitation solution，by dissociating with dissociation dissociation． The concentration and purity of the supernatant and the two dissociation solutions were determined respectively． Result The optimum condition for the albumin complexation was pH 8. 5，at room temperature for 2 h，and 2. 3% rivanol，optimum condition for gamma globulin． complexation was pH 6. 0，at room temperature for 2 h，and 2. 3% rivanol． The optimum condition for the albumin，dissociation was 0. 6% sodium chloride，0. 25% sodium caprylate and pH 1. 37，and optimum condition for the the gamma globulin dissociation was 0. 85% sodium chloride，1. 3% glycine，and pH 6. 8. Conclusion The dog albumin and gamma globulin could be successfully separated and purified with rivanol method．

Key words: Rivanol method, complexation and dissociation, dog plasma separation and purification

中图分类号:

R392. 7

隋丽华, 陈旖, 李瑞生, 刘宏伟, 赵彦斌, 张小飞, 黄永莉, 刘红燕, 刘永梅, 叶华虎. 利凡诺方法分离纯化犬血浆蛋白络合及解离条件探讨[J]. 实验动物科学, 2012, 29(06): 30-33.

SUI Li-Hua, Chen-Yi, LI Rui-Sheng, LIU Hong-Wei, ZHAO Yan-Bin, Zhang-Xiao-Fei, HUANG Yong-Li, LIU Hong-Yan, Liu-Yong-Mei, Ye-Hua-Hu. Investigation on Conditions of Complexation and Dissociation in Separating and Purifying Dog Plasma Proteins with Rivanol Method[J]. laboratory animal science, 2012, 29(06): 30-33.