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Creating hybrid proteins by insertion of exogenous peptides into permissive sites of a class A β‐lactamase
Author(s) -
Ruth Nadia,
Quinting Birgit,
Mainil Jacques,
Hallet Bernard,
Frère JeanMarie,
Huygen Kris,
Galleni Moreno
Publication year - 2008
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2008.06646.x
Subject(s) - heterologous , epitope , v3 loop , peptide , escherichia coli , enterotoxin , biology , antibody , peptide sequence , protein subunit , microbiology and biotechnology , chemistry , biochemistry , genetics , gene
Insertion of heterologous peptide sequences into a protein carrier may impose structural constraints that could help the peptide to adopt a proper fold. This concept could be the starting point for the development of a new generation of safe subunit vaccines based on the expression of poorly immunogenic epitopes. In the present study, we characterized the tolerance of the TEM‐1 class A β‐lactamase to the insertion of two different peptides, the V3 loop of the gp120 protein of HIV, and the thermostable STa enterotoxin produced by enterotoxic Escherichia coli . Insertion of the V3 loop of the HIV gp120 protein into the TEM‐1 scaffold yielded insoluble and poorly produced proteins. By contrast, four hybrid β‐lactamases carrying the STa peptide were efficiently produced and purified. Immunization of BALB/c mice with these hybrid proteins produced high levels of TEM‐1‐specific antibodies, together with significant levels of neutralizing antibodies against STa.