Produção e caracterização da porção Fab do anticorpo anti-digoxina utilizando a tecnologia de phage display. / Production and characterization of the Fab portion of anti-digoxin antibody by phage display technology.

AUTOR(ES)

FONTE

DATA DE PUBLICAÇÃO

27/03/2012

RESUMO

Digoxin is a pharmaceutical used in the control of cardiac dysfunction. Its therapeutic window is very narrow. To counteract the toxic effect, polyclonal anti-digoxin Fab fragments are commercially available. Our goal was to obtain variants of monoclonal anti-digoxin Fab fragments by phage display technology, which allows the generation of high affinity and specificity antibody fragments. Anti-digoxin Fab fragments combinatorial library was constructed into pComb3X vector from total RNA of anti-digoxin hybridoma. Six clones were isolated and the heavy chain presented the same sequence. For the light chain, 2 clones were identical, one was a pseudogene and other one presented a distinct amino acid in the CDR2. Four clones presenting variations in the framework 1 were induced to express soluble Fab fragments, all positive for anti-digoxin binding in ELISA assays and Western blotting. The specific binding of the antibody was further confirmed by BIAcore, which allowed ranking of the clones.

ASSUNTO(S)

anticorpos monoclonais cardiovascular system clonagem cloning digoxin digoxina drugs fármacos hibridomas hybridomas monoclonal antibodies sistema cardiovascular

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