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标题: 从噬菌体展示的人合成Fab文库中产生和表征来自乙型肝炎病 [打印本页]

作者: StephenW    时间: 2018-10-13 13:41     标题: 从噬菌体展示的人合成Fab文库中产生和表征来自乙型肝炎病

J Microbiol Biotechnol. 2018 Aug 28;28(8):1376-1383. doi: 10.4014/jmb.1803.03056.
Generation and Characterization of a Neutralizing Human Monoclonal Antibody to Hepatitis B Virus PreS1 from a Phage-Displayed Human Synthetic Fab Library.
Jo G1, Jeong MS1, Wi J2, Kim DH3, Kim S1, Kim D1, Yoon JY1, Chae H1, Kim KH3,4, Hong HJ1,2.
Author information

1
    Department of Systems Immunology, College of Biomedical Science, Kangwon National University, Chuncheon 24341, Republic of Korea.
2
    Scripps Korea Antibody Institute, Chuncheon 24341, Republic of Korea.
3
    Department of Pharmacology, Center for Cancer Research and Diagnostic Medicine, IBST, School of Medicine, Konkuk University, Seoul 05029, Republic of Korea.
4
    Research Institute of Medical Sciences, Konkuk University, Seoul 05029, Republic of Korea.

Abstract

The hepatitis B virus (HBV) envelope contains small (S), middle (M), and large (L) proteins. PreS1 of the L protein contains a receptor-binding motif crucial for HBV infection. This motif is highly conserved among 10 HBV genotypes (A-J), making it a potential target for the prevention of HBV infection. In this study, we successfully generated a neutralizing human monoclonal antibody (mAb), 1A8 (IgG1), that recognizes the receptor-binding motif of preS1 using a phage-displayed human synthetic Fab library. Analysis of the antigen-binding activity of 1A8 for different genotypes indicated that it can specifically bind to the preS1 of major HBV genotypes (A-D). Based on Bio-Layer interferometry, the affinity (KD) of 1A8 for the preS1 of genotype C was 3.55 nM. 1A8 immunoprecipitated the hepatitis B virions of genotypes C and D. In an in vitro neutralization assay using HepG2 cells overexpressing the cellular receptor sodium taurocholate cotransporting polypeptide, 1A8 effectively neutralized HBV infection with genotype D. Taken together, the results suggest that 1A8 may neutralize the four HBV genotypes. Considering that genotypes A-D are most prevalent, 1A8 may be a neutralizing human mAb with promising potential in the prevention and treatment of HBV infection.
KEYWORDS:

Hepatitis B virus; human monoclonal antibody; neutralizing antibody; phage display; preS1; synthetic antibody library

PMID:
    30301315
DOI:
    10.4014/jmb.1803.03056
作者: StephenW    时间: 2018-10-13 13:41

J Microbiol Biotechnol。 2018年8月28日; 28(8):1376-1383。 doi:10.4014 / jmb.1803.03056。
从噬菌体展示的人合成Fab文库中产生和表征来自乙型肝炎病毒PreS1的中和人单克隆抗体。
Jo G1,Jeong MS1,Wi J2,Kim DH3,Kim S1,Kim D1,Yoon JY1,Chae H1,Kim KH3,4,Hong HJ1,2。
作者信息

1
    江原大学生物医学科学系系统免疫学系,大韩民国春川24341。
2
    Scripps Korea Antibody Institute,Chuncheon 24341,大韩民国。
3
    韩国首尔康氧大学医学院IBST癌症研究与诊断医学中心药理学系,首尔05029。
4
    韩国Konkuk大学医学科学研究所,首尔05029。

抽象

乙型肝炎病毒(HBV)包膜含有小(S),中(M)和大(L)蛋白。 L蛋白的PreS1含有对HBV感染至关重要的受体结合基序。该基序在10种HBV基因型(A-J)中高度保守,使其成为预防HBV感染的潜在靶标。在这项研究中,我们成功地产生了中和人单克隆抗体(mAb)1A8(IgG1),其使用噬菌体展示的人合成Fab文库识别preS1的受体结合基序。对不同基因型的1A8的抗原结合活性的分析表明它可以特异性结合主要HBV基因型(A-D)的preS1。基于生物层干涉测定法,1A8对基因型C的preS1的亲和力(KD)为3.55nM。 1A8免疫沉淀基因型C和D的乙型肝炎病毒粒子。在体外中和试验中,使用过表达细胞受体牛磺胆酸钠协同转运多肽的HepG2细胞,1A8有效地中和了基因型D的HBV感染。总之,结果表明1A8可以中和四种HBV基因型。考虑到基因型A-D是最普遍的,1A8可以是中和人mAb,具有预防和治疗HBV感染的潜力。
关键词:

乙型肝炎病毒;人单克隆抗体;中和抗体;噬菌体展示;前S1;合成抗体库

结论:
    30301315
DOI:
    10.4014 / jmb.1803.03056




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