Extended plasma half-life of albumin-binding domain fused human IgA upon pH-dependent albumin engagement of human FcRn in vitro and in vivo
Mester, Simone; Evers, Mitchell; Meyer, Saskia; Nilsen, Jeannette; Greiff, Victor; Sandlie, Inger; Leusen, Jeanette; Andersen, Jan Terje
(2021) mAbs, volume 13, issue 1, pp. 1 - 14
(Article)
Abstract
Albumin has a serum half-life of 3 weeks in humans. This feature can be used to improve the pharmacokinetics of shorter-lived biologics. For instance, an albumin-binding domain (ABD) can be used to recruit albumin. A prerequisite for such design is that the ABD-albumin interaction does not interfere with pH-dependent binding
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of albumin to the human neonatal Fc receptor (FcRn), as FcRn acts as the principal regulator of the half-life of albumin. Thus, there is a need to know how ABDs act in the context of fusion partners and human FcRn. Here, we studied the binding and transport properties of human immunoglobulin A1 (IgA1), fused to a Streptococcus protein G-derived engineered ABD, in in vitro and in vivo systems harboring human FcRn. IgA has great potential as a therapeutic protein, but its short half-life is a major drawback. We demonstrate that ABD-fused IgA1 binds human FcRn pH-dependently and is rescued from cellular degradation in a receptor-specific manner in the presence of albumin. This occurs when ABD is fused to either the light or the heavy chain. In human FcRn transgenic mice, IgA1-ABD in complex with human albumin, gave 4-6-fold extended half-life compared to unmodified IgA1, where the light chain fusion showed the longest half-life. When the heavy chain-fused protein was pre-incubated with an engineered human albumin with improved FcRn binding, cellular rescue and half-life was further enhanced. Our study reveals how an ABD, which does not interfere with albumin binding to human FcRn, may be used to extend the half-life of IgA.Abbreviations: ABD - Albumin binding domain, ADA - anti-drug-antibodies, ADCC - Antibody-dependent cellular cytotoxicity, ELISA - Enzyme-linked Immunosorbent assay, FcαRI - Fcα receptor, FcγR - Fcγ receptor, FcRn - The neonatal Fc receptor, GST - Glutathione S-transferase, HC - Heavy chain, HERA - Human endothelial cell-based recycling assay, Her2 - Human epidermal growth factor 2, HMEC - Human microvascular endothelial cells, IgG - Immunoglobulin G, IgA - Immunoglobulin A, LC - Light chain, QMP - E505Q/T527M/K573P, WT - Wild type.
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Keywords: albumin-binding-domain (ABD), half-life, human FcRn transgenic mice, human serum albumin (HSA), Immunoglobulin A (IgA), the neonatal Fc receptor (FcRn), Immunology and Allergy, Immunology, Research Support, Non-U.S. Gov't, Journal Article
ISSN: 1942-0862
Publisher: Landes Bioscience
Note: Funding Information: We are grateful to Sathiaruby Sivaganesh and Ignacio Navas Camacho for excellent technical assistance, and we thank Dr. Wayne I. Lencer (Boston Children’s Hospital, Harvard Medical School and Harvard Digestive Diseases center) for the HMEC1 cell line stably expressing HA-hFcRn-EGFP. Funding: This work was supported in part by the Research Council of Norway through its Center of Excellence funding scheme (project no. 179573). J.T.A. and J.N. were supported by the Research Council of Norway (Grant no. 230526 and 287927), and J.T.A. and S.M. by South-Eastern Norway Regional Health Authority (Grant no. 40018). M.E. was funded by KWF (Grant no. 7650). V.G. was supported by a UiO World-Leading Research Community grant, the UiO:LifeSciences Convergence Environment Immunolingo, EU Horizon 2020 iReceptorplus (#825821), and Research Council of Norway FRIPRO project (#300740). Funding Information: This work was supported by the Horizon 2020 Framework Programme [825821]; KWF Kankerbestrijding [7650]; Norges Forskningsråd [230526]; Norges Forskningsråd [179573]; Norges Forskningsråd [300740]; South-Eastern Norway Regional Health Authority [40018]; Research Council of Norway [179573, 230526, 287927]; South-Eastern Norway Regional Health Authority [40018]. We are grateful to Sathiaruby Sivaganesh and Ignacio Navas Camacho for excellent technical assistance, and we thank Dr. Wayne I. Lencer (Boston Children’s Hospital, Harvard Medical School and Harvard Digestive Diseases center) for the HMEC1 cell line stably expressing HA-hFcRn-EGFP. Funding: This work was supported in part by the Research Council of Norway through its Center of Excellence funding scheme (project no. 179573). J.T.A. and J.N. were supported by the Research Council of Norway (Grant no. 230526 and 287927), and J.T.A. and S.M. by South-Eastern Norway Regional Health Authority (Grant no. 40018). M.E. was funded by KWF (Grant no. 7650). V.G. was supported by a UiO World-Leading Research Community grant, the UiO:LifeSciences Convergence Environment Immunolingo, EU Horizon 2020 iReceptorplus (#825821), and Research Council of Norway FRIPRO project (#300740). Publisher Copyright: © 2021 The Author(s). Published with license by Taylor & Francis Group, LLC.
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