Isolation of human anti-Bothrops atrox venom antibodies from a naive library displayed on filamentous phage surface (2006)
- Authors:
- Autor USP: KIPNIS, THEREZA LIBERMAN - ICB
- Unidade: ICB
- Assunto: IMUNOLOGIA
- Language: Inglês
- Abstract: Introduction and Objectives: The utilization of antibody fragment libraries displayed on filamentous phage surface is an interesting technique to obtain antibodies against different kind of antigens in a fast and safe way. In this work, human monoclonal antibody reactive to Bothrops atrox venom crude extract were selected from a human Fab library previously reported (Dantas-Barbosa et al., Gen. Mol. Res. 4(2): 126-140 - 2005). The aim of this work is to obtain a panel of human antibodies against different B. atrox venom components that can be used in the future in passive immunotherapy. Methods and Results: The 1.45 x 108 Fab functional form library was used to perform six selection rounds. The Fabs were selected by their ability to bind to B. atrox venom crude extract adsorbed on a microtiter plate. In each round the phages were washed out and eluted. The best enrichment of the binding population was achieved from round 2 to 3.Randomly chosen clones from these rounds were genetic and phenotypically analyzed. The phagemids were cleaved by Sfi I and the Fab 1,6 Kb corresponding fragments were identified in most of the clones. Recombinant Fab fusion phages from 46 different clones were produced in E. coli supernatants inoculated in one mL culture broth in deep well plates. The resulting supernatants were used in a Dot Blot assay with anti-M13 antibody to evaluate phage production. The phages were also used in a Phage ELISA assay to detectvenom crude extract affinity. Seventeen positive clones diversity was assessed by BstO I fingerprinting. From these, eight Fab coding regions were sequenced. Conclusion: The data shown here suggest that human Fabs against Bothrops atrox venom were selected from a combinatorial phage displayed library. These results prove the potential of this library as a human antibody source against different antigens. The isolated human antibodies can also contribute to low immunogenic antiofidic sera formulations.
- Imprenta:
- Source:
- Título: Abstracts
- Conference titles: Meeting of the Brazilian Society for Immunology
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ABNT
CAFÉ-MENDES, C C et al. Isolation of human anti-Bothrops atrox venom antibodies from a naive library displayed on filamentous phage surface. 2006, Anais.. São Paulo: Instituto de Ciências Biomédicas, Universidade de São Paulo, 2006. . Acesso em: 28 dez. 2025. -
APA
Café-Mendes, C. C., Albuquerque, F. C., Kanashiro, M. M., Kipnis, T. L., Brígido, M. M., & Maranhão, A. Q. (2006). Isolation of human anti-Bothrops atrox venom antibodies from a naive library displayed on filamentous phage surface. In Abstracts. São Paulo: Instituto de Ciências Biomédicas, Universidade de São Paulo. -
NLM
Café-Mendes CC, Albuquerque FC, Kanashiro MM, Kipnis TL, Brígido MM, Maranhão AQ. Isolation of human anti-Bothrops atrox venom antibodies from a naive library displayed on filamentous phage surface. Abstracts. 2006 ;[citado 2025 dez. 28 ] -
Vancouver
Café-Mendes CC, Albuquerque FC, Kanashiro MM, Kipnis TL, Brígido MM, Maranhão AQ. Isolation of human anti-Bothrops atrox venom antibodies from a naive library displayed on filamentous phage surface. Abstracts. 2006 ;[citado 2025 dez. 28 ] - Study of the peritrophic matrix protein composition in aedes aegypti
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