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Design of a Peptibody Consisting of the Antimicrobial Peptide dhvar5 and a llama Variable Heavy‐chain Antibody Fragment
Author(s) -
Szynol A.,
De Haard J. J. W.,
Veerman E. C.,
De Soet J. J.,
Van Nieuw Amerongen A. V.
Publication year - 2006
Publication title -
chemical biology and drug design
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.59
H-Index - 77
eISSN - 1747-0285
pISSN - 1747-0277
DOI - 10.1111/j.1747-0285.2006.00395.x
Subject(s) - immunoconjugate , peptide , recombinant dna , escherichia coli , antimicrobial , fusion protein , chemistry , antimicrobial peptides , biochemistry , biology , microbiology and biotechnology , antibody , monoclonal antibody , gene , immunology
Immunoconjugates have been widely studied as potential therapeutics for infectious diseases to direct unspecific antimicrobials to pathogens. In this study, the recombinant approach was used for expression of the immunoconjugate composed of the variable domain of a llama heavy‐chain antibody (VHH) against Streptococcus mutans and dhvar5, a synthetic antimicrobial peptide. Before cloning, the impact of the elongation of the peptide termini on its biological activity was evaluated by chemical synthesis of the N‐ or C‐termini extended dhvar5 peptides. As the elongation of the C‐terminus had a greater influence on decline of the antimicrobial activity, the N‐terminal fusion was designed. To promote in vivo release of the active peptide, a factor Xa cleavage site was inserted between VHH and dhvar5. Propagation of transformed Escherichia coli with the constructed plasmid was only possible in the absence of isopropyl β‐D‐thiogalactoside (IPTG). Although these data demonstrate that the diminished antimicrobial activity of dhvar5 by the N‐terminal fusion to VHH was not sufficient for the protection of the bacterial host cells against the peptide lethal effect, an insight into propeptides biological activities may be beneficial not only for new and more successful rearrangement of the VHH–dhvar5 immunoconjugate construct, but also design of the other recombinant molecules composed of peptides toxic to host cells.