| 摘要 |
The filamentous bacteriophage is a common immunogenic carrier for generating anti-peptide antibodies against recombinant peptides displayed on its surface. For immunizations, peptides are displayed as fusions to either the minor coat protein, pIII, or, more commonly the major coat protein, pVIII. Phage displaying a poorly expressed malarial peptide on pVII I were compared to phage bearing the same peptide on pIII , and to phage that were chemically coupled to a synthetic version of the peptide using the crosslinker, sulfo-succinimidyl 4-(N-maleimido-methyl)cyclohexane-1-carboxylate (sulfo-SMCC) for their abili ty to induce an anti-peptide Ab response in mice. We also included in this comparison the same peptide displayed as a recombinant fusion to the maltose-binding protein (MBP) of Escherichia coli, as well as the synthetic counterpart as chemically crosslinked to MBP with sulfo-SMCC. The results demonstrate that only the chemically-crosslinked conjugates elicited a stron g anti-peptide antibody response by ELISA, probably due to the higher number of peptides th at were displayed by the phage and MBP conjugates as compared to their recombinant counterpart s. The amount of peptide that was covalently linked to the phage was increased approximate ly two-fold when a lysine residue was engineered near the N-terminus of mature pVIII. The ease of chemically conjugating synthetic peptides to phage, and of modifying pVIII by genetic engineering, suggests new roles for the filamentous phage in vaccine studies.
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