Binding of a therapeutic protein to a long-circulating plasma protein can result in a strongly extended half-life. Among these plasma proteins, albumin and immunoglobulins are of special interest because of their exceptionally long half-life, which is to a great extent determined by recycling through the neonatal Fc receptor (FcRn). Many strategies have been established employing reversible binding to albumin, e.g. using an albumin-binding domain from streptococcal protein G. We show here that the half-life of a recombinant antibody molecule can also be prolonged by fusion to a single immunoglobulin-binding domain (IgBD) from staphylococcal protein A. This domain (domain B, SpAB) is composed of 56 amino acid residues and was fused to the C-terminus of a bispecific single-chain diabody (scDb). The scDb-SpAB fusion protein was produced in HEK293 cells and retained its antigen-binding activity as shown by enzyme-linked immunosorbent assay and flow cytometry. Furthermore, the fusion protein was capable of binding to human and mouse IgG in a pH-dependent manner. In mice, the terminal half-life of the fusion protein was improved from ∼1–2 h of the unmodified scDb to 11.8 h. Although the fusion protein did not reach the long half-life seen for IgG, our results established the applicability of a single bacterial IgBD for half-life extension purposes.
%0 Journal Article
%1 Unverdorben2012
%A Unverdorben, Felix
%A Färber-Schwarz, Aline
%A Richter, Fabian
%A Hutt, Meike
%A Kontermann, Roland E.
%D 2012
%J Protein Engineering, Design and Selection
%K 2012 izi kontermann
%N 2
%P 81--88
%R 10.1093/protein/gzr061
%T Half-life extension of a single-chain diabody by fusion to domain B of staphylococcal protein A
%U http://www.ncbi.nlm.nih.gov/pubmed/22238430
%V 25
%X Binding of a therapeutic protein to a long-circulating plasma protein can result in a strongly extended half-life. Among these plasma proteins, albumin and immunoglobulins are of special interest because of their exceptionally long half-life, which is to a great extent determined by recycling through the neonatal Fc receptor (FcRn). Many strategies have been established employing reversible binding to albumin, e.g. using an albumin-binding domain from streptococcal protein G. We show here that the half-life of a recombinant antibody molecule can also be prolonged by fusion to a single immunoglobulin-binding domain (IgBD) from staphylococcal protein A. This domain (domain B, SpAB) is composed of 56 amino acid residues and was fused to the C-terminus of a bispecific single-chain diabody (scDb). The scDb-SpAB fusion protein was produced in HEK293 cells and retained its antigen-binding activity as shown by enzyme-linked immunosorbent assay and flow cytometry. Furthermore, the fusion protein was capable of binding to human and mouse IgG in a pH-dependent manner. In mice, the terminal half-life of the fusion protein was improved from ∼1–2 h of the unmodified scDb to 11.8 h. Although the fusion protein did not reach the long half-life seen for IgG, our results established the applicability of a single bacterial IgBD for half-life extension purposes.
%@ 1741-0134 (Electronic)$\backslash$r1741-0126 (Linking)
@article{Unverdorben2012,
abstract = {Binding of a therapeutic protein to a long-circulating plasma protein can result in a strongly extended half-life. Among these plasma proteins, albumin and immunoglobulins are of special interest because of their exceptionally long half-life, which is to a great extent determined by recycling through the neonatal Fc receptor (FcRn). Many strategies have been established employing reversible binding to albumin, e.g. using an albumin-binding domain from streptococcal protein G. We show here that the half-life of a recombinant antibody molecule can also be prolonged by fusion to a single immunoglobulin-binding domain (IgBD) from staphylococcal protein A. This domain (domain B, SpAB) is composed of 56 amino acid residues and was fused to the C-terminus of a bispecific single-chain diabody (scDb). The scDb-SpAB fusion protein was produced in HEK293 cells and retained its antigen-binding activity as shown by enzyme-linked immunosorbent assay and flow cytometry. Furthermore, the fusion protein was capable of binding to human and mouse IgG in a pH-dependent manner. In mice, the terminal half-life of the fusion protein was improved from ∼1–2 h of the unmodified scDb to 11.8 h. Although the fusion protein did not reach the long half-life seen for IgG, our results established the applicability of a single bacterial IgBD for half-life extension purposes.},
added-at = {2018-02-01T16:21:30.000+0100},
author = {Unverdorben, Felix and F{\"{a}}rber-Schwarz, Aline and Richter, Fabian and Hutt, Meike and Kontermann, Roland E.},
biburl = {https://puma.ub.uni-stuttgart.de/bibtex/218fa10f37ead25fe06eecf90b2f52d95/cristiano},
doi = {10.1093/protein/gzr061},
interhash = {f7fa7674ee6d57892f81bb5f203cc771},
intrahash = {18fa10f37ead25fe06eecf90b2f52d95},
isbn = {1741-0134 (Electronic)$\backslash$r1741-0126 (Linking)},
issn = {17410126},
journal = {Protein Engineering, Design and Selection},
keywords = {2012 izi kontermann},
month = feb,
number = 2,
pages = {81--88},
pmid = {22238430},
timestamp = {2019-01-17T13:18:41.000+0100},
title = {{Half-life extension of a single-chain diabody by fusion to domain B of staphylococcal protein A}},
url = {http://www.ncbi.nlm.nih.gov/pubmed/22238430},
volume = 25,
year = 2012
}
