Cloning of human ScFv against digoxin from a semi-synthetic phage antibody library
- VernacularTitle:从半合成噬菌体抗体库中克隆抗地高辛人单链抗体
- Author:
Dexin LIU
;
Xiaoying LI
;
Bing HUA
;
Al ET
- Publication Type:Journal Article
- Keywords:
Phage antibody library Human ScFv Digoxin Toxication
- From:
Chinese Journal of Immunology
2000;0(11):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To clone human anti digoxin ScFv from a human semi synthetic phage antibody library.Methods:①Digoxin BSA conjugate(Dig BSA) was preparaed by a modified periodate oxidation method.②A semi synthetic phage antibody library was panned against immobilized Dig BSA.Collected clones were analysed by ELISA,inhibition ELISA and DNA Sequencing.Results:①During the four rounds panning against Dig BSA,the enrichment of the eluted phage particles was observed;②Analysis of the eluted clones identified one clone that could bind Dig as well as other digitalis.③Sequencing analysis showed that the variable genes of the positive clone belonged to VH5 and V?1 subgroup respectively.Conclusion:Human Dig specific ScFv that could bind Dig and other digitalis has been cloned from a semi synthetic phage antibody library which may provide a potential reagent for the diagnosis and therapy of Dig toxication.
