Immunogenic epitope scanning in bacteriolytic enzymes Pal and Cpl-1 and engineering Pal to escape antibody responses
Bacteriolytic enzymes are promising antibacterial agents, but they can cause a typical immune response in vivo. In this study, we used a targeted modification method for two antibacterial endolysins, Pal and Cpl-1. We identified the key immunogenic amino acids, and designed and tested new, bacteriol...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-09-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1075774/full |
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| author | Marek Adam Harhala Marek Adam Harhala Katarzyna Gembara Katarzyna Gembara Izabela Rybicka Zuzanna Maria Kaźmierczak Zuzanna Maria Kaźmierczak Paulina Miernikiewicz Joanna Marta Majewska Wiktoria Budziar Anna Nasulewicz-Goldeman Daniel C. Nelson Barbara Owczarek Krystyna Dąbrowska Krystyna Dąbrowska |
| author_facet | Marek Adam Harhala Marek Adam Harhala Katarzyna Gembara Katarzyna Gembara Izabela Rybicka Zuzanna Maria Kaźmierczak Zuzanna Maria Kaźmierczak Paulina Miernikiewicz Joanna Marta Majewska Wiktoria Budziar Anna Nasulewicz-Goldeman Daniel C. Nelson Barbara Owczarek Krystyna Dąbrowska Krystyna Dąbrowska |
| author_sort | Marek Adam Harhala |
| collection | DOAJ |
| description | Bacteriolytic enzymes are promising antibacterial agents, but they can cause a typical immune response in vivo. In this study, we used a targeted modification method for two antibacterial endolysins, Pal and Cpl-1. We identified the key immunogenic amino acids, and designed and tested new, bacteriolytic variants with altered immunogenicity. One new variant of Pal (257-259 MKS → TFG) demonstrated decreased immunogenicity while a similar mutant (257-259 MKS → TFK) demonstrated increased immunogenicity. A third variant (280-282 DKP → GGA) demonstrated significantly increased antibacterial activity and it was not cross-neutralized by antibodies induced by the wild-type enzyme. We propose this variant as a new engineered endolysin with increased antibacterial activity that is capable of escaping cross-neutralization by antibodies induced by wild-type Pal. We show that efficient antibacterial enzymes that avoid cross-neutralization by IgG can be developed by epitope scanning, in silico design, and substitutions of identified key amino acids with a high rate of success. Importantly, this universal approach can be applied to many proteins beyond endolysins and has the potential for design of numerous biological drugs. |
| first_indexed | 2024-03-12T00:36:19Z |
| format | Article |
| id | doaj.art-921cc27f8a7645c29fa84e3ab39311ad |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-03-12T00:36:19Z |
| publishDate | 2023-09-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-921cc27f8a7645c29fa84e3ab39311ad2023-09-15T09:35:50ZengFrontiers Media S.A.Frontiers in Immunology1664-32242023-09-011410.3389/fimmu.2023.10757741075774Immunogenic epitope scanning in bacteriolytic enzymes Pal and Cpl-1 and engineering Pal to escape antibody responsesMarek Adam Harhala0Marek Adam Harhala1Katarzyna Gembara2Katarzyna Gembara3Izabela Rybicka4Zuzanna Maria Kaźmierczak5Zuzanna Maria Kaźmierczak6Paulina Miernikiewicz7Joanna Marta Majewska8Wiktoria Budziar9Anna Nasulewicz-Goldeman10Daniel C. Nelson11Barbara Owczarek12Krystyna Dąbrowska13Krystyna Dąbrowska14Laboratory of Phage Molecular Biology, Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw, PolandResearch and Development Centre, Regional Specialist Hospital, Wroclaw, PolandLaboratory of Phage Molecular Biology, Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw, PolandResearch and Development Centre, Regional Specialist Hospital, Wroclaw, PolandLaboratory of Phage Molecular Biology, Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw, PolandLaboratory of Phage Molecular Biology, Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw, PolandResearch and Development Centre, Regional Specialist Hospital, Wroclaw, PolandLaboratory of Phage Molecular Biology, Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw, PolandLaboratory of Phage Molecular Biology, Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw, PolandResearch and Development Centre, Regional Specialist Hospital, Wroclaw, PolandLaboratory of Phage Molecular Biology, Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw, PolandInstitute for Bioscience and Biotechnology Research, University of Maryland, Rockville, MD, United StatesLaboratory of Phage Molecular Biology, Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw, PolandLaboratory of Phage Molecular Biology, Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw, PolandResearch and Development Centre, Regional Specialist Hospital, Wroclaw, PolandBacteriolytic enzymes are promising antibacterial agents, but they can cause a typical immune response in vivo. In this study, we used a targeted modification method for two antibacterial endolysins, Pal and Cpl-1. We identified the key immunogenic amino acids, and designed and tested new, bacteriolytic variants with altered immunogenicity. One new variant of Pal (257-259 MKS → TFG) demonstrated decreased immunogenicity while a similar mutant (257-259 MKS → TFK) demonstrated increased immunogenicity. A third variant (280-282 DKP → GGA) demonstrated significantly increased antibacterial activity and it was not cross-neutralized by antibodies induced by the wild-type enzyme. We propose this variant as a new engineered endolysin with increased antibacterial activity that is capable of escaping cross-neutralization by antibodies induced by wild-type Pal. We show that efficient antibacterial enzymes that avoid cross-neutralization by IgG can be developed by epitope scanning, in silico design, and substitutions of identified key amino acids with a high rate of success. Importantly, this universal approach can be applied to many proteins beyond endolysins and has the potential for design of numerous biological drugs.https://www.frontiersin.org/articles/10.3389/fimmu.2023.1075774/fulldeimmunizationendolysinPALCpl-1epitope engineeringVirScan |
| spellingShingle | Marek Adam Harhala Marek Adam Harhala Katarzyna Gembara Katarzyna Gembara Izabela Rybicka Zuzanna Maria Kaźmierczak Zuzanna Maria Kaźmierczak Paulina Miernikiewicz Joanna Marta Majewska Wiktoria Budziar Anna Nasulewicz-Goldeman Daniel C. Nelson Barbara Owczarek Krystyna Dąbrowska Krystyna Dąbrowska Immunogenic epitope scanning in bacteriolytic enzymes Pal and Cpl-1 and engineering Pal to escape antibody responses Frontiers in Immunology deimmunization endolysin PAL Cpl-1 epitope engineering VirScan |
| title | Immunogenic epitope scanning in bacteriolytic enzymes Pal and Cpl-1 and engineering Pal to escape antibody responses |
| title_full | Immunogenic epitope scanning in bacteriolytic enzymes Pal and Cpl-1 and engineering Pal to escape antibody responses |
| title_fullStr | Immunogenic epitope scanning in bacteriolytic enzymes Pal and Cpl-1 and engineering Pal to escape antibody responses |
| title_full_unstemmed | Immunogenic epitope scanning in bacteriolytic enzymes Pal and Cpl-1 and engineering Pal to escape antibody responses |
| title_short | Immunogenic epitope scanning in bacteriolytic enzymes Pal and Cpl-1 and engineering Pal to escape antibody responses |
| title_sort | immunogenic epitope scanning in bacteriolytic enzymes pal and cpl 1 and engineering pal to escape antibody responses |
| topic | deimmunization endolysin PAL Cpl-1 epitope engineering VirScan |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1075774/full |
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