Fusion of M13 bacteriophage pVIII protein with antigen protein using SpyTag/SpyCatcher system to increase antibody titer in rainbow trout (Oncorhynchus mykiss)
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M13 bacteriophage is a virus that specifically infects Escherichia coli exhibiting an F pilus. Its structure comprises five different coat proteins surrounding its genome. Since the development of phage display technology, which enables the display of foreign peptides/proteins on phage surfaces, various applications from medicine to nanomaterials have utilized phages. Notably, studies using phages as delivery tools for drugs and protective antigens have garnered significant attention. The delivery of foreign antigens via M13 phage display limits the size of the molecule expressed on the major coat protein (pVIII) to 15 amino acids. In this study, by applying the SpyTag/SpyCatcher system, we displayed a foreign antigen that cannot be displayed on the pVIII with the existing system. The conjugation reaction between SpyTag-displaying phage on the pVIII protein and SpyCatcher-fused protein was confirmed by SDS-PAGE and Western blot. In the immunization experiment in rainbow trout (Oncorhynchus mykiss), the group administered the foreign protein displaying phage showed a higher antibody titer than the group administered the foreign protein alone. These results suggest that large antigens can be displayed on the pVIII protein of M13 phage using SpyTag/SpyCatcher system. Furthermore, unlike subunit vaccines requiring adjuvants to induce a sufficient immune re- sponse, the phage can act as a self-adjuvanting agent.
Abstract
M13 bacteriophage is a virus that specifically infects Escherichia coli exhibiting an F pilus. Its structure comprises five different coat proteins surrounding its genome. Since the development of phage display technology, which enables the display of foreign peptides/proteins on phage surfaces, various applications from medicine to nanomaterials have utilized phages. Notably, studies using phages as delivery tools for drugs and protective antigens have garnered significant attention. The delivery of foreign antigens via M13 phage display limits the size of the molecule expressed on the major coat protein (pVIII) to 15 amino acids. In this study, by applying the SpyTag/SpyCatcher system, we displayed a foreign antigen that cannot be displayed on the pVIII with the existing system. The conjugation reaction between SpyTag-displaying phage on the pVIII protein and SpyCatcher-fused protein was confirmed by SDS-PAGE and Western blot. In the immunization experiment in rainbow trout (Oncorhynchus mykiss), the group administered the foreign protein displaying phage showed a higher antibody titer than the group administered the foreign protein alone. These results suggest that large antigens can be displayed on the pVIII protein of M13 phage using SpyTag/SpyCatcher system. Furthermore, unlike subunit vaccines requiring adjuvants to induce a sufficient immune re- sponse, the phage can act as a self-adjuvanting agent.
참고문헌 (18)
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