Screening and identification of human monoclonal antibodies against low-calcium response V antigen of Yersinia pestis
10.3969/j.issn.1002-2694.2024.00.014
- VernacularTitle:鼠疫耶尔森菌低钙应答V抗原人源单克隆抗体筛选与鉴定
- Author:
Li ZHANG
1
,
2
;
Bin-Yang ZHENG
;
Qi ZHANG
;
Hai-Lian WU
;
Hong-Xin PAN
;
Feng-Cai ZHU
;
Hai-Sheng WU
;
Jian-Fang ZHOU
Author Information
1. 中国疾病预防控制中心病毒病预防控制所,传染病溯源预警和智能决策全国重点实验室,北京 102206
2. 江苏省疾病预防控制中心,南京 210009
- Keywords:
Yersinia pestis;
low-calcium response V antigen;
phage display;
recombinant antibody;
protective effect
- From:
Chinese Journal of Zoonoses
2024;40(1):15-20
- CountryChina
- Language:Chinese
-
Abstract:
To characterize human antibodies against low-calcium response V(LcrV)antigen of Yersinia pestis,the mono-clonal antibodies were screened and assayed.Antibody gene was derived from peripheral blood mononuclear cells of the vaccin-ees immunized by plague subunit vaccine in phase Ⅱb clinical trial.Human ScFv antibody library was constructed by phage dis-play.After panning library by using recombinant LcrV antigen,antibody variable genes were sequenced and converted into IgG1 format to evaluate its binding specificity and relevant parameters.An anti-plague human ScFv antibody library was estab-lished contained 7.54× 108 independent clones.After panning by LcrV antigen,3 human antibodies named as RV-B4,RV-D1 and RV-E8,respectively,were identified.Using indirect enzyme-linked immunosorbent assay(ELISA)and Western blot(WB),the specific bindings of the mAbs to LcrV antigen were confirmed.The dissociation constant(KD)of them to LcrV is 2.1 nmol/L,1.24 nmol/L and 42 nmol/L,respectively.Minor protective efficacy was found among 3 human antibodies in Y.pestis 141-infected mice.Three anti-LcrV monoclonal antibodies generated from immunized vaccinees were binding specific antibod-ies and could not block plague infection in mice.These antibodies are the potential candidate reagents for basic research of plague immunity and the application of plague diagnosis.
