Affinity maturation of a single-chain antibody against rabies virus
10.3760/cma.j.issn.1003-9279.2016.03.018
- VernacularTitle:易错PCR提高人源抗狂犬病毒单链抗体的亲和力
- Author:
Lina SUN
1
;
Chuan LI
;
Yang LIU
;
Dexin LI
;
Mifang LIANG
Author Information
1. 中国疾病预防控制中心病毒病预防控制所
- Keywords:
Rabies virus;
Single-chain antibody;
Affinity
- From:
Chinese Journal of Experimental and Clinical Virology
2016;30(3):328-332
- CountryChina
- Language:Chinese
-
Abstract:
Objective To obtain a human single-chain antibody against rabies virus (RV) with high affinity by error-prone PCR technology.Methods A combinatorial scFv antibody mutant library was constructed by error-prone PCR amplifying VH and VL of scFv RV08.After package by hyperphage,the scFv phage mutant library was panned by ELISA and IFA with purified RV AG.The selected mutant antibodies were converted to full human IgG antibodies with the VH and VK Express cassettes.The affinity of mutant antibodies was measured by non-competitive enzyme immunoassay.Results The mutant antibody library with capacity of 6 × 106 cfu/ml contained target gene insertion rate of 100％.Heavy chain and light chain were sequenced with mutation frequency of about 0.95％ and 1.2％ respectively.Finally 5 mutant antibodies with high affinity were obtained,of which HL46,HL2 and HL37 had higher affinity with 1.79 times,1.47 times and 1.23 times of RV08 respectively.Conclusions In this study,error-prone PCR technology and phage display technology combination enables to obtain significantly improved affinity of antibody.
