Humanization of an anti-Bothrops atrox phospholipase 'A IND.2'SCFV antibody fragment (2006)
- USP affiliated authors: KIPNIS, THEREZA LIBERMAN - ICB
- Unidades: ICB
- Subjects: IMUNOLOGIA
- Language: Inglês
- Abstract: Introduction and Objectives: Until now, the serotherapy is the only treatment to poisonous animal accidents, despite of the toxicity driven by the equine antibody utilization. Nowadays several studies have been developed to minimize these collateral effects. The Phospholipase A2 (PLA2) is a major antigen found in the Bothrops atrox venom and causes miotoxicity, inflammation and necrosis. Monoclonal antibodies (mAbs) against this antigen were produced in mice and were able to neutralize the miotoxic activity of PLA2 efficiently (Biochem Pharmacol. 64(7):1179-86). The objective of this work is to develop a humanized form of one of these anti-PLA2 antibodies, aiming its clinical use for snake poisoning treatment. Methods and Results: Initially the light and heavy chains of the mAb were determined by cloning Ig cDNA from mAb´s hybridoma. The anti-PLA2 antibody heavy and light chains are coded by VH 36-60 and Vk 21-2 gene fragments, respectively.To humanize this antibody, the light and heavy chain's CDRs of the murine mAb were transferred to a human framework, found on the database of human germline V genes. The closest framework chosen were a human VH4/JH6 for the heavy and a B3/Jk4 for the light variable domain, used for the original murine CDR introduction. The proposed Fv has 16 non-human residues in the light chain and 9 in the heavy chain. The sequence was synthesized as a scFv (single chain variablefragment) and cloned in expression vectors for Escherichia coli and Pichia pastoris heterologous expressions. Conclusion: The sequence of the anti-PLA2 antibody was determined and used to identify the closest human variable gene for both light and heavy domain. This analysis permitted the humanized scFv proposal. The proposed sequence maintains a minimal fraction of murine residues (10%). The recombinant Fv antibody fragment will now be compared with the original antibody for its affinity towards PLA2 and neutralization proprieties.
- Título do periódico: Abstracts
- Conference title: Meeting of the Brazilian Society for Immunology
ABNTPIMENTEL, B M S; KANASHIRO, M M; ALBUQUERQUE, F C; et al. Humanization of an anti-Bothrops atrox phospholipase 'A IND.2'SCFV antibody fragment. Anais.. São Paulo: [s.n.], 2006.
APAPimentel, B. M. S., Kanashiro, M. M., Albuquerque, F. C., Kipnis, T. L., Maranhão, A. Q., & Brígido, M. M. (2006). Humanization of an anti-Bothrops atrox phospholipase 'A IND.2'SCFV antibody fragment. In Abstracts. São Paulo.
NLMPimentel BMS, Kanashiro MM, Albuquerque FC, Kipnis TL, Maranhão AQ, Brígido MM. Humanization of an anti-Bothrops atrox phospholipase 'A IND.2'SCFV antibody fragment. Abstracts. 2006 ;
VancouverPimentel BMS, Kanashiro MM, Albuquerque FC, Kipnis TL, Maranhão AQ, Brígido MM. Humanization of an anti-Bothrops atrox phospholipase 'A IND.2'SCFV antibody fragment. Abstracts. 2006 ;
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