Enhanced plasma half-life and efficacy of engineered human albumin-fused GLP-1 despite enzymatic cleavage of its C-terminal end
Abstract Albumin has a long plasma half-life due to engagement of the neonatal Fc receptor (FcRn), which prevents intracellular degradation. However, its C-terminal end can be cleaved by carboxypeptidase A, and removal of the last leucine residue (L585) weakens receptor binding, reducing its half-li...
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Nature Portfolio
2025-05-01
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| Series: | Communications Biology |
| Online Access: | https://doi.org/10.1038/s42003-025-08249-8 |
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| author | Jeannette Nilsen Kristin Hovden Aaen Sopisa Benjakul Fulgencio Ruso-Julve Thomas Uwe Greiner Daniela Bejan Maria Stensland Sachin Singh Tilman Schlothauer Inger Sandlie Jan Terje Andersen |
| author_facet | Jeannette Nilsen Kristin Hovden Aaen Sopisa Benjakul Fulgencio Ruso-Julve Thomas Uwe Greiner Daniela Bejan Maria Stensland Sachin Singh Tilman Schlothauer Inger Sandlie Jan Terje Andersen |
| author_sort | Jeannette Nilsen |
| collection | DOAJ |
| description | Abstract Albumin has a long plasma half-life due to engagement of the neonatal Fc receptor (FcRn), which prevents intracellular degradation. However, its C-terminal end can be cleaved by carboxypeptidase A, and removal of the last leucine residue (L585) weakens receptor binding, reducing its half-life from 20 days to 3.5 days in humans. This biology has so far been overlooked when designing human albumin-fused biologics. Thus, there is a need for an engineering strategy to secure favorable FcRn binding and pharmacokinetic properties. Here, we show that a branched aliphatic amino acid or methionine at position 585 of albumin is required for optimal receptor binding, which cannot be replaced to prevent enzymatic cleavage without negatively affecting FcRn engagement. As a solution, we report that C-terminally cleaved albumin can be efficiently rescued from intracellular degradation by introducing amino acid substitutions that improve FcRn binding. This albumin-engineering strategy was also effective when applied with a therapeutic fusion partner, glucagon-like peptide 1 (GLP-1), resulting in a 2-fold increase in plasma half-life and prolonged efficacy in human FcRn transgenic mice. We demonstrate how human albumin fusions should be tailored to ensure a long plasma half-life and enhanced efficacy of fused biologics, despite potential C-terminal cleavage in vivo. |
| format | Article |
| id | doaj-art-d22d3339131a40e8b331fa1611c1b4bf |
| institution | DOAJ |
| issn | 2399-3642 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Communications Biology |
| spelling | doaj-art-d22d3339131a40e8b331fa1611c1b4bf2025-08-20T03:22:12ZengNature PortfolioCommunications Biology2399-36422025-05-018111710.1038/s42003-025-08249-8Enhanced plasma half-life and efficacy of engineered human albumin-fused GLP-1 despite enzymatic cleavage of its C-terminal endJeannette Nilsen0Kristin Hovden Aaen1Sopisa Benjakul2Fulgencio Ruso-Julve3Thomas Uwe Greiner4Daniela Bejan5Maria Stensland6Sachin Singh7Tilman Schlothauer8Inger Sandlie9Jan Terje Andersen10Department of Immunology, Oslo University Hospital RikshospitaletDepartment of Immunology, Oslo University Hospital RikshospitaletDepartment of Immunology, Oslo University Hospital RikshospitaletDepartment of Immunology, Oslo University Hospital RikshospitaletWallenberg Laboratory, Department of Molecular and Clinical Medicine, Institute of Medicine, University of GothenburgDepartment of Immunology, Oslo University Hospital RikshospitaletDepartment of Immunology, Oslo University Hospital RikshospitaletDepartment of Immunology, Oslo University Hospital RikshospitaletRoche Pharma Research and Early Development (pRED), Therapeutic Modalities, Roche Innovation Center Munich, Roche Diagnostics GmbHDepartment of Biosciences, University of OsloDepartment of Immunology, Oslo University Hospital RikshospitaletAbstract Albumin has a long plasma half-life due to engagement of the neonatal Fc receptor (FcRn), which prevents intracellular degradation. However, its C-terminal end can be cleaved by carboxypeptidase A, and removal of the last leucine residue (L585) weakens receptor binding, reducing its half-life from 20 days to 3.5 days in humans. This biology has so far been overlooked when designing human albumin-fused biologics. Thus, there is a need for an engineering strategy to secure favorable FcRn binding and pharmacokinetic properties. Here, we show that a branched aliphatic amino acid or methionine at position 585 of albumin is required for optimal receptor binding, which cannot be replaced to prevent enzymatic cleavage without negatively affecting FcRn engagement. As a solution, we report that C-terminally cleaved albumin can be efficiently rescued from intracellular degradation by introducing amino acid substitutions that improve FcRn binding. This albumin-engineering strategy was also effective when applied with a therapeutic fusion partner, glucagon-like peptide 1 (GLP-1), resulting in a 2-fold increase in plasma half-life and prolonged efficacy in human FcRn transgenic mice. We demonstrate how human albumin fusions should be tailored to ensure a long plasma half-life and enhanced efficacy of fused biologics, despite potential C-terminal cleavage in vivo.https://doi.org/10.1038/s42003-025-08249-8 |
| spellingShingle | Jeannette Nilsen Kristin Hovden Aaen Sopisa Benjakul Fulgencio Ruso-Julve Thomas Uwe Greiner Daniela Bejan Maria Stensland Sachin Singh Tilman Schlothauer Inger Sandlie Jan Terje Andersen Enhanced plasma half-life and efficacy of engineered human albumin-fused GLP-1 despite enzymatic cleavage of its C-terminal end Communications Biology |
| title | Enhanced plasma half-life and efficacy of engineered human albumin-fused GLP-1 despite enzymatic cleavage of its C-terminal end |
| title_full | Enhanced plasma half-life and efficacy of engineered human albumin-fused GLP-1 despite enzymatic cleavage of its C-terminal end |
| title_fullStr | Enhanced plasma half-life and efficacy of engineered human albumin-fused GLP-1 despite enzymatic cleavage of its C-terminal end |
| title_full_unstemmed | Enhanced plasma half-life and efficacy of engineered human albumin-fused GLP-1 despite enzymatic cleavage of its C-terminal end |
| title_short | Enhanced plasma half-life and efficacy of engineered human albumin-fused GLP-1 despite enzymatic cleavage of its C-terminal end |
| title_sort | enhanced plasma half life and efficacy of engineered human albumin fused glp 1 despite enzymatic cleavage of its c terminal end |
| url | https://doi.org/10.1038/s42003-025-08249-8 |
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