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- Title
A strategy for the generation of specific human antibodies by directed evolution and phage display.
- Authors
Riaño-Umbarila, Lidia; Juárez-González, Victor Rivelino; Olamendi-Portugal, Timoteo; Ortíz-León, Mauricio; Possani, Lourival Domingos; Becerril, Baltazar
- Abstract
This study describes the construction of a library of single-chain antibody fragments (scFvs) from a single human donor by individual amplification of all heavy and light variable domains (1.1 × 108 recombinants). The library was panned using the phage display technique, which allowed selection of specific scFvs (3F and C1) capable of recognizing Cn2, the major toxic component ofCentruroides noxiusscorpion venom. The scFv 3F was maturedin vitroby three cycles of directed evolution. The use of stringent conditions in the third cycle allowed the selection of several improved clones. The best scFv obtained (6009F) was improved in terms of its affinity by 446-fold, from 183 nm(3F) to 410 pm. This scFv 6009F was able to neutralize 2 LD50 of Cn2 toxin when a 1 : 10 molar ratio of toxin-to-antibody fragment was used. It was also able to neutralize 2 LD50 of the whole venom. These results pave the way for the future generation of recombinant human antivenoms.
- Subjects
IMMUNOGLOBULINS; CENTRUROIDES; BUTHIDAE; TOXINS; ANTIVENINS; VENOM
- Publication
FEBS Journal, 2005, Vol 272, Issue 10, p2591
- ISSN
1742-464X
- Publication type
Article
- DOI
10.1111/j.1742-4658.2005.04687.x