Half-life extension of efficiently produced DARPin serum albumin fusions as a function of FcRn affinity and recycling.

Merten, Hannes; Brandl, Fabian; Zimmermann, Martina; Schaefer, Jonas V; Irpinio, Linda; Sand, Kine M K; Nilsen, Jeannette; Andersen, Jan Terje; Zangemeister-Wittke, Uwe; Plückthun, Andreas (2021). Half-life extension of efficiently produced DARPin serum albumin fusions as a function of FcRn affinity and recycling. European journal of pharmaceutics and biopharmaceutics, 167, pp. 104-113. Elsevier 10.1016/j.ejpb.2021.07.011

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Serum albumin shows slow clearance from circulation due to neonatal Fc receptor (FcRn)-mediated recycling and has been used for half-life extension. We report here fusions to a high-affinity DARPin, binding to Epithelial Cell Adhesion Molecule (EpCAM). We developed a novel, efficient expression system for such fusion proteins in Pichia pastoris with titers above 300 mg/L of lab-scale shake-flask culture. Since human serum albumin (HSA) does not bind to the murine FcRn, half-lives of therapeutic candidates are frequently measured in human FcRn transgenic mice, limiting useable tumor models. Additionally, serum albumins with extended half-life have been designed. We tested HSA7, motivated by its previously claimed extraordinarily long half-life in mice, which we could not confirm. Instead, we determined a half-life of only 29 h for HSA7, comparable to MSA. The fusion of HSA7 to a DARPin showed a similar half-life. To rationalize these findings, we measured binding kinetics and affinities to murine and human FcRn. Briefly, HSA7 showed affinity to murine FcRn only in the micromolar range, comparable to MSA to its cognate murine FcRn, and an affinity in the nanomolar range only to the human FcRn. This explains the comparable half-life of MSA and HSA7 in mice, while wild-type-HSA has a half-life of only 21 h, as it does not bind the murine FcRn and is not recycled. Thus, HSA-fusions with improved FcRn-affinity, such as HSA7, can be used for preclinical experiments in mice when FcRn transgenes cannot be used, as they reflect better the complex FcRn-mediated recycling and distribution mechanisms.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Pre-clinic Human Medicine > Institute of Pharmacology

UniBE Contributor:

Zangemeister-Wittke, Uwe

Subjects:

600 Technology > 610 Medicine & health

ISSN:

0939-6411

Publisher:

Elsevier

Language:

English

Submitter:

Celine Joray

Date Deposited:

11 Nov 2021 09:24

Last Modified:

11 Nov 2021 09:24

Publisher DOI:

10.1016/j.ejpb.2021.07.011

PubMed ID:

34303832

Uncontrolled Keywords:

Binding affinity DARPin FcRn HSA7 Half-life Pichia pastoris Serum albumin

BORIS DOI:

10.48350/160300

URI:

https://boris.unibe.ch/id/eprint/160300

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