The University of Southampton
University of Southampton Institutional Repository

Increasing the serum persistence of an IgG fragment by random mutagenesis

Increasing the serum persistence of an IgG fragment by random mutagenesis
Increasing the serum persistence of an IgG fragment by random mutagenesis

The major histocompatibility complex (MHC) class l-related receptor FcRn is involved in regulating serum gammaglobulin (IgG) levels in mice. With the aim of increasing the serum half-life of a recombinant murine Fcγ1 fragment, the affinity for binding to FcRn at pH 6,0 has been increased by random mutagenesis of Thr252, Thr254, and Thr256 followed by selection using bacteriophage display. These residues were chosen as they are in proximity to the FcRn-IgG (Fc) interaction site. Two mutants with higher affinity (due to lower off-rates) than the wild-type Fc have been isolated and analyzed in pharmacokinetic studies in mice. The mutant with the highest affinity has a significantly longer serum half-life than the wild type fragment, despite its lower off-rate from FcRn at pH 7.4. The results provide support for the involvement of FcRn in the home-ostasis of serum IgGs in mice. The indications that a homologous FcRn regulates IgG levels in humans suggest that this approach has implications for increasing the serum persistence of therapeutic antibodies.

Antibody engineering, Phage display, Pharmacokinetics, Surface plasmon resonance
1087-0156
637-640
Ghetie, Victor
b7b50946-2bd9-4896-b841-6bbfba8237c2
Popov, Serguei
84ac1427-131d-4f26-a1ca-88d3589607e0
Borvak, Jozef
0756de83-6795-49ab-9df5-07e3dc3ae05e
Radu, Caius
cf50ccf6-c03c-4d8f-abd8-56681251f990
Matesoi, Diana
0ed9f055-aaa2-4d84-a478-f45852460980
Medesan, Corneliu
0de1d9e4-c8d9-4896-bcca-63f8b24e8f2c
Ober, Raimund J.
31f4d47f-fb49-44f5-8ff6-87fc4aff3d36
Ward, E. Sally
b31c0877-8abe-485f-b800-244a9d3cd6cc
Ghetie, Victor
b7b50946-2bd9-4896-b841-6bbfba8237c2
Popov, Serguei
84ac1427-131d-4f26-a1ca-88d3589607e0
Borvak, Jozef
0756de83-6795-49ab-9df5-07e3dc3ae05e
Radu, Caius
cf50ccf6-c03c-4d8f-abd8-56681251f990
Matesoi, Diana
0ed9f055-aaa2-4d84-a478-f45852460980
Medesan, Corneliu
0de1d9e4-c8d9-4896-bcca-63f8b24e8f2c
Ober, Raimund J.
31f4d47f-fb49-44f5-8ff6-87fc4aff3d36
Ward, E. Sally
b31c0877-8abe-485f-b800-244a9d3cd6cc

Ghetie, Victor, Popov, Serguei, Borvak, Jozef, Radu, Caius, Matesoi, Diana, Medesan, Corneliu, Ober, Raimund J. and Ward, E. Sally (1997) Increasing the serum persistence of an IgG fragment by random mutagenesis. Nature Biotechnology, 15 (7), 637-640. (doi:10.1038/nbt0797-637).

Record type: Article

Abstract

The major histocompatibility complex (MHC) class l-related receptor FcRn is involved in regulating serum gammaglobulin (IgG) levels in mice. With the aim of increasing the serum half-life of a recombinant murine Fcγ1 fragment, the affinity for binding to FcRn at pH 6,0 has been increased by random mutagenesis of Thr252, Thr254, and Thr256 followed by selection using bacteriophage display. These residues were chosen as they are in proximity to the FcRn-IgG (Fc) interaction site. Two mutants with higher affinity (due to lower off-rates) than the wild-type Fc have been isolated and analyzed in pharmacokinetic studies in mice. The mutant with the highest affinity has a significantly longer serum half-life than the wild type fragment, despite its lower off-rate from FcRn at pH 7.4. The results provide support for the involvement of FcRn in the home-ostasis of serum IgGs in mice. The indications that a homologous FcRn regulates IgG levels in humans suggest that this approach has implications for increasing the serum persistence of therapeutic antibodies.

Full text not available from this repository.

More information

Accepted/In Press date: 30 April 1997
Published date: 1 July 1997
Keywords: Antibody engineering, Phage display, Pharmacokinetics, Surface plasmon resonance

Identifiers

Local EPrints ID: 425013
URI: https://eprints.soton.ac.uk/id/eprint/425013
ISSN: 1087-0156
PURE UUID: 23040f56-ac89-4a81-82c3-ff59ef389f8f
ORCID for Raimund J. Ober: ORCID iD orcid.org/0000-0002-1290-7430
ORCID for E. Sally Ward: ORCID iD orcid.org/0000-0003-3232-7238

Catalogue record

Date deposited: 09 Oct 2018 16:30
Last modified: 20 Jul 2019 00:22

Export record

Altmetrics

Contributors

Author: Victor Ghetie
Author: Serguei Popov
Author: Jozef Borvak
Author: Caius Radu
Author: Diana Matesoi
Author: Corneliu Medesan
Author: Raimund J. Ober ORCID iD
Author: E. Sally Ward ORCID iD

University divisions

Download statistics

Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.

View more statistics

Atom RSS 1.0 RSS 2.0

Contact ePrints Soton: eprints@soton.ac.uk

ePrints Soton supports OAI 2.0 with a base URL of https://eprints.soton.ac.uk/cgi/oai2

This repository has been built using EPrints software, developed at the University of Southampton, but available to everyone to use.

We use cookies to ensure that we give you the best experience on our website. If you continue without changing your settings, we will assume that you are happy to receive cookies on the University of Southampton website.

×