Genetic and immunological comparison of anti-botulinum type A antibodies from immune and non-immune human phage libraries

被引:80
作者
Amersdorfer, P
Wong, C
Smith, T
Chen, S
Deshpande, S
Sheridan, R
Marks, JD
机构
[1] Univ Calif San Francisco, San Francisco Gen Hosp, Dept Anesthesia & Pharmaceut Chem, San Francisco, CA 94110 USA
[2] USA, Med Res Inst Infect Dis, Toxinol Div, Frederick, MD 21702 USA
[3] USA, Med Res Inst Chem Def, Neurotoxicol Branch, Div Pathophysiol, Aberdeen Proving Ground, MD 21010 USA
关键词
botulinum; antibody engineering; vaccine; phage display;
D O I
10.1016/S0264-410X(01)00482-0
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Understanding the antibody response in botulinum intoxication is important for vaccine design and passive prophylaxis. To investigate this activity, we have studied the immune response to BoNT/A (botulinum neurotoxin serotype A) binding domain (H-C) at the molecular level using phage display. The scFv antibodies were isolated from V gene repertoires prepared from (a) human volunteer immunized with pentavalent botulinum toxoid and (b) non-immune human peripheral blood lymphocytes and spleenocytes. A large panel of serotype specific phage expressing botulinum binding scFv could be selected from both libraries. Epitope mapping of immune scFv binders towards BoNT/A HC revealed surprisingly a limited number of scFv recognizing conformational epitopes that corresponded to two distinct groups, clusters I and II Only scFv from cluster I exhibited neutralizing activity in the mouse hemidiaphragm assay. Anti- BoNT/A HC clones derived from a non-immune library could be conveniently grouped into clusters III-XI and appeared to share no overlapping epitopes with cluster I or H. In addition they showed no neutralization of toxin at biologically significant concentrations. We therefore suggest that a vaccine based on the pentavalent botulinum toxoid directs the humoral immune response to a limited number of immunodominant epitopes exposed on the binding domain H-C. (C) 2002 Elsevier Science Ltd. All rights reserved.
引用
收藏
页码:1640 / 1648
页数:9
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