Improved Biodistribution and Extended Serum Half-Life of a Bacteriophage Endolysin by Albumin Binding Domain Fusion
The increasing number of multidrug-resistant bacteria intensifies the need to develop new antimicrobial agents. Endolysins are bacteriophage-derived enzymes that degrade the bacterial cell wall and hold promise as a new class of highly specific and versatile antimicrobials. One major limitation to t...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2018-11-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fmicb.2018.02927/full |
| _version_ | 1818527285641216000 |
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| author | Johan Seijsing Anna M. Sobieraj Nadia Keller Yang Shen Annelies S. Zinkernagel Martin J. Loessner Mathias Schmelcher |
| author_facet | Johan Seijsing Anna M. Sobieraj Nadia Keller Yang Shen Annelies S. Zinkernagel Martin J. Loessner Mathias Schmelcher |
| author_sort | Johan Seijsing |
| collection | DOAJ |
| description | The increasing number of multidrug-resistant bacteria intensifies the need to develop new antimicrobial agents. Endolysins are bacteriophage-derived enzymes that degrade the bacterial cell wall and hold promise as a new class of highly specific and versatile antimicrobials. One major limitation to the therapeutic use of endolysins is their often short serum circulation half-life, mostly due to kidney excretion and lysosomal degradation. One strategy to increase the half-life of protein drugs is fusion to the albumin-binding domain (ABD). By high-affinity binding to serum albumin, ABD creates a complex with large hydrodynamic volume, reducing kidney excretion and lysosomal degradation. The aim of this study was to investigate the in vitro antibacterial activity and in vivo biodistribution and half-life of an engineered variant of the Staphylococcus aureus phage endolysin LysK. The ABD sequence was introduced at different positions within the enzyme, and lytic activity of each variant was determined in vitro and ex vivo in human serum. Half-life and biodistribution were assessed in vivo by intravenous injection of europium-labeled proteins into C57BL/6 wild-type mice. Our data demonstrates that fusion of the endolysin to ABD improves its serum circulation half-life and reduces its deposition in the kidneys in vivo. The most active construct reduced S. aureus counts in human serum ex vivo by 3 logs within 60 min. We conclude that ABD fusions provide an effective strategy to extend the half-life of antibacterial enzymes, supporting their therapeutic potential for treatment of systemic bacterial infections. |
| first_indexed | 2024-12-11T06:34:09Z |
| format | Article |
| id | doaj.art-7208ba3a8031415ca4ff598cb3fc9bc9 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-11T06:34:09Z |
| publishDate | 2018-11-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-7208ba3a8031415ca4ff598cb3fc9bc92022-12-22T01:17:26ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2018-11-01910.3389/fmicb.2018.02927429454Improved Biodistribution and Extended Serum Half-Life of a Bacteriophage Endolysin by Albumin Binding Domain FusionJohan Seijsing0Anna M. Sobieraj1Nadia Keller2Yang Shen3Annelies S. Zinkernagel4Martin J. Loessner5Mathias Schmelcher6Laboratory of Food Microbiology, Institute of Food, Nutrition and Health, ETH Zürich, Zürich, SwitzerlandLaboratory of Food Microbiology, Institute of Food, Nutrition and Health, ETH Zürich, Zürich, SwitzerlandDivision of Infectious Diseases and Hospital Epidemiology, University Hospital Zürich – University of Zürich, Zürich, SwitzerlandLaboratory of Food Microbiology, Institute of Food, Nutrition and Health, ETH Zürich, Zürich, SwitzerlandDivision of Infectious Diseases and Hospital Epidemiology, University Hospital Zürich – University of Zürich, Zürich, SwitzerlandLaboratory of Food Microbiology, Institute of Food, Nutrition and Health, ETH Zürich, Zürich, SwitzerlandLaboratory of Food Microbiology, Institute of Food, Nutrition and Health, ETH Zürich, Zürich, SwitzerlandThe increasing number of multidrug-resistant bacteria intensifies the need to develop new antimicrobial agents. Endolysins are bacteriophage-derived enzymes that degrade the bacterial cell wall and hold promise as a new class of highly specific and versatile antimicrobials. One major limitation to the therapeutic use of endolysins is their often short serum circulation half-life, mostly due to kidney excretion and lysosomal degradation. One strategy to increase the half-life of protein drugs is fusion to the albumin-binding domain (ABD). By high-affinity binding to serum albumin, ABD creates a complex with large hydrodynamic volume, reducing kidney excretion and lysosomal degradation. The aim of this study was to investigate the in vitro antibacterial activity and in vivo biodistribution and half-life of an engineered variant of the Staphylococcus aureus phage endolysin LysK. The ABD sequence was introduced at different positions within the enzyme, and lytic activity of each variant was determined in vitro and ex vivo in human serum. Half-life and biodistribution were assessed in vivo by intravenous injection of europium-labeled proteins into C57BL/6 wild-type mice. Our data demonstrates that fusion of the endolysin to ABD improves its serum circulation half-life and reduces its deposition in the kidneys in vivo. The most active construct reduced S. aureus counts in human serum ex vivo by 3 logs within 60 min. We conclude that ABD fusions provide an effective strategy to extend the half-life of antibacterial enzymes, supporting their therapeutic potential for treatment of systemic bacterial infections.https://www.frontiersin.org/article/10.3389/fmicb.2018.02927/fullstaphylococciantibiotic alternativesbiodistributionhalf-lifeendolysinalbumin binding domain (ABD) |
| spellingShingle | Johan Seijsing Anna M. Sobieraj Nadia Keller Yang Shen Annelies S. Zinkernagel Martin J. Loessner Mathias Schmelcher Improved Biodistribution and Extended Serum Half-Life of a Bacteriophage Endolysin by Albumin Binding Domain Fusion Frontiers in Microbiology staphylococci antibiotic alternatives biodistribution half-life endolysin albumin binding domain (ABD) |
| title | Improved Biodistribution and Extended Serum Half-Life of a Bacteriophage Endolysin by Albumin Binding Domain Fusion |
| title_full | Improved Biodistribution and Extended Serum Half-Life of a Bacteriophage Endolysin by Albumin Binding Domain Fusion |
| title_fullStr | Improved Biodistribution and Extended Serum Half-Life of a Bacteriophage Endolysin by Albumin Binding Domain Fusion |
| title_full_unstemmed | Improved Biodistribution and Extended Serum Half-Life of a Bacteriophage Endolysin by Albumin Binding Domain Fusion |
| title_short | Improved Biodistribution and Extended Serum Half-Life of a Bacteriophage Endolysin by Albumin Binding Domain Fusion |
| title_sort | improved biodistribution and extended serum half life of a bacteriophage endolysin by albumin binding domain fusion |
| topic | staphylococci antibiotic alternatives biodistribution half-life endolysin albumin binding domain (ABD) |
| url | https://www.frontiersin.org/article/10.3389/fmicb.2018.02927/full |
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