A Three Monoclonal Antibody Combination Potently Neutralizes Multiple Botulinum Neurotoxin Serotype E Subtypes

被引：35

作者：

Garcia-Rodriguez, Consuelo ^{[1
]}

Razai, Ali ^{[1
,2
]}

Geren, Isin N. ^{[1
,3
]}

Lou, Jianlong ^{[1
]}

Conrad, Fraser ^{[1
]}

Wen, Wei-Hua ^{[1
]}

Farr-Jones, Shauna ^{[1
]}

Smith, Theresa J. ^{[4
]}

Brown, Jennifer L. ^{[5
]}

Skerry, Janet C. ^{[5
]}

Smith, Leonard A. ^{[6
]}

Marks, James D. ^{[1
]}

机构：

[1] Univ Calif San Francisco, Zuckerberg San Francisco Gen Hosp & Trauma Ctr, Dept Anesthesia & Perioperat Care, Room 3C-38,1001 Potrero Ave, San Francisco, CA 94110 USA

[2] Knobbe Martin, 2040 Main St,14th Floor, Irvine, CA 92614 USA

[3] Istanbul Medeniyet Univ, Dept Mol Biol & Genet, Fac Sci, Unalan Mahallesi,Unalan Sokak,D100 Karayolu, TR-34700 Uskudar Istanbul, Turkey

[4] USAMRIID, Mol & Translat Sci Div, Ft Detrick, MD 21702 USA

[5] USAMRIID, Keaki Technol LLC, Ft Detrick, MD 21702 USA

[6] USAMRIID, Med Countermeasures Technol, Ft Detrick, MD 21702 USA

来源：

TOXINS | 2018年 / 10卷 / 03期

关键词：

botulinum neurotoxin; oligoclonal antibodies; serotype E botulism; recombinant antibodies; antibody engineering; mouse neutralization assay; botulinum antitoxin; DOMAIN-BASED ASSAYS; CLOSTRIDIUM-BOTULINUM; CRYSTAL-STRUCTURE; CHAIN ANTIBODIES; E STRAINS; BINDING; BUTYRICUM; AFFINITY; GENE; IDENTIFICATION;

D O I：

10.3390/toxins10030105

中图分类号：

TS2 [食品工业];

学科分类号：

0832 ;

摘要：

Human botulism is most commonly caused by botulinum neurotoxin (BoNT) serotypes A, B, and E. For this work, we sought to develop a human monoclonal antibody (mAb)-based antitoxin capable of binding and neutralizing multiple subtypes of BoNT/E. Libraries of yeast-displayed single chain Fv (scFv) antibodies were created from the heavy and light chain variable region genes of humans immunized with pentavalent-toxoid- and BoNT/E-binding scFv isolated by Fluorescence-Activated Cell Sorting (FACS). A total of 10 scFv were isolated that bound one or more BoNT/E subtypes with nanomolar-level equilibrium dissociation constants (K-D). By diversifying the V-regions of the lead mAbs and selecting for cross-reactivity, we generated three scFv that bound all four BoNT/E subtypes tested at three non-overlapping epitopes. The scFvs were converted to IgG that had K-D values for the different BoNT/E subtypes ranging from 9.7 nM to 2.28 pM. An equimolar combination of the three mAbs was able to potently neutralize BoNT/E1, BoNT/E3, and BoNT/E4 in a mouse neutralization assay. The mAbs have potential utility as therapeutics and as diagnostics capable of recognizing multiple BoNT/E subtypes. A derivative of the three-antibody combination (NTM-1633) is in pre-clinical development with an investigational new drug (IND) application filing expected in 2018.

引用

页数：16

共 50 条

[1] Neutralization of Botulinum Neurotoxin by a Human Monoclonal Antibody Specific for the Catalytic Light Chain [J].