Preparation of conotoxin GI antiserum and determination of neutralization activity
10.7644/j.issn.1674-9960.2017.05.003
- VernacularTitle:芋螺毒素GI抗血清制备及中和活性研究
- Author:
Yifei TANG
;
Xiaobo HU
;
Shuo YU
;
Qiuyun DAI
- Keywords:
conotoxins;
bovine serum albumin;
coupling;
immunization;
neutralization activity
- From:Military Medical Sciences
2017;41(5):338-341
- CountryChina
- Language:Chinese
-
Abstract:
Objective To determine whether the antiserum produced by immunizing mice with conotoxin GI coupled with bovine serum albumin (BSA) could neutralize GI conotoxin.Methods The GI-BSA was prepared by glutaraldehyde-coupled method,and the mice were immunized with the GI-BSA to produce antiserum.The antibody neutralization assay was used to test the detoxication of the antiserum.Results The SDS-PAGE protein electrophoresis showed that the coupling reaction of GI hapten with BSA was successful.The two distinct protein bands of GI-BSA were more than 120×103.Each mouse was immunized four times with 99 μg every two weeks.After the fourth immunization,the serum neutralization titer was more than 1:64 000.After the intraperitoneal injection of the mixture of 100 or 200 μl of the antiserum and different doses of GI,75% of the mice survived in the group with 100 μl of the antiserum and 1× LD50 GI(16.3 μg/kg).The same percentage of mice also survived in the group of with 200 μl of serum and 25.8 μg/kg of GI.Conclusion The antiserum produced by immunizing mice with GI-BSA exhibits significant detoxication activity to conotoxin GI.
