A recombinant Fab neutralizes dengue virus in vitro

被引:43
作者
Thullier, P
Lafaye, P
Mégret, F
Deubel, V
Jouan, A
Mazié, JC
机构
[1] Inst Pasteur, Lab Ingn Anticorps, F-75724 Paris 15, France
[2] Serv Sante Armees, Ctr Rech, Dept Biol Agents Transmissibles, La Tronche, France
[3] Inst Pasteur, Unite Arbovirus & Virus Fievres Hemorrag, Paris, France
关键词
Fab; recombinant; dengue; neutralization; phages;
D O I
10.1016/S0168-1656(99)00037-1
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
A recombinant Fab that recognizes a neutralizing epitope located in the (296-400) region of protein E of dengue virus was obtained from cloned hybridoma cells secreting the mouse monoclonal antibody (mAb) 3E11. The Fd and light chain antibody genes were amplified by polymerase chain reaction, cloned into the phagemid vector pMad, expressed in bacteria to produce Fab fragments and sequenced. The mAb 4E11, in particular its light chain complementary-determining regions, shared homologies with two other anti-viral mAbs. The affinity of the parental mAb and the cloned Fab to the MalE-E(296-400) fusion protein were shown to be of the same magnitude, i.e. nanomolar. Fab 4E11 neutralization capacity was found between 8 and 4-times or less lower than that of mAb 4E11, depending on serotypes, thus the Fab could have a smaller antiviral activity than the mAb in vitro. (C) 1999 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:183 / 190
页数:8
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