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 m...

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Bibliographic Details
Published in:Nature biotechnology 1997-07, Vol.15 (7), p.637-640
Main Authors: Ghetie, Victor, Popov, Serguei, Borvak, Jozef, Radu, Caius, Matesoi, Diana, Medesan, Corneliu, Ober, Raimund J, Ward, E. Sally
Format: Article
Language:English
Subjects:
Agriculture
Animals
Base Sequence
Bioinformatics
Biological and medical sciences
Biomedical and Life Sciences
Biomedical Engineering/Biotechnology
Biomedicine
Biotechnology
Fundamental and applied biological sciences. Psychology
Half-Life
Humans
Hydrogen-Ion Concentration
Immunoglobulin Fragments - blood
Immunoglobulin Fragments - genetics
Immunoglobulin G - blood
Immunoglobulin G - genetics
Life Sciences
Methods. Procedures. Technologies
Mice
Mutagenesis
Oligodeoxyribonucleotides - genetics
Protein Engineering
Receptors, IgG - genetics
Receptors, IgG - metabolism
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Summary: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.
ISSN:1087-0156
1546-1696
DOI:10.1038/nbt0797-637