Cross-species analysis of Fc engineered anti-Lewis-Y human IgG1 variants in human neonatal receptor transgenic mice reveal importance of S254 and Y436 in binding human neonatal Fc receptor

Ingrid J.G. Burvenich, William Farrugia, Fook T. Lee, Bruno Catimel, Zhanqi Liu, Dahna Makris, Diana Cao, Graeme J. O'Keefe, Martin W. Brechbiel, Dylan King, Violeta Spirkoska, Laura C. Allan, Paul A. Ramsland, Andrew M. Scott

Research output: Contribution to journalArticleResearchpeer-review

3 Citations (Scopus)

Abstract

IgG has a long half-life through engagement of its Fc region with the neonatal Fc receptor (FcRn). The FcRn binding site on IgG1 has been shown to contain I253 and H310 in the CH2 domain and H435 in the CH3 domain. Altering the half-life of IgG has been pursued with the aim to prolong or reduce the half-life of therapeutic IgGs. More recent studies have shown that IgGs bind differently to mouse and human FcRn. In this study we characterize a set of hu3S193 IgG1 variants with mutations in the FcRn binding site. A double mutation in the binding site is necessary to abrogate binding to murine FcRn, whereas a single mutation in the FcRn binding site is sufficient to no longer detect binding to human FcRn and create hu3S193 IgG1 variants with a half-life similar to previously studied hu3S193 F(ab’)2 (t1/2β, I253A, 12.23 h; H310A, 12.94; H435A, 12.57; F(ab’)2, 12.6 h). Alanine substitutions in S254 in the CH2 domain and Y436 in the CH3 domain showed reduced binding in vitro to human FcRn and reduced elimination half-lives in huFcRn transgenic mice (t1/2β, S254A, 37.43 h; Y436A, 39.53 h; wild-type, 83.15 h). These variants had minimal effect on half-life in BALB/c nu/nu mice (t1/2β, S254A, 119.9 h; Y436A, 162.1 h; wild-type, 163.1 h). These results provide insight into the interaction of human Fc by human FcRn, and are important for antibody-based therapeutics with optimal pharmacokinetics for payload strategies used in the clinic.

Original languageEnglish
Pages (from-to)775-786
Number of pages12
JournalmAbs
Volume8
Issue number4
DOIs
Publication statusPublished - 18 May 2016

Keywords

  • Antibody engineering
  • Fc receptors
  • Molecular biology
  • Neonatal Fc receptor
  • Transgenic mice

Cite this

Burvenich, Ingrid J.G. ; Farrugia, William ; Lee, Fook T. ; Catimel, Bruno ; Liu, Zhanqi ; Makris, Dahna ; Cao, Diana ; O'Keefe, Graeme J. ; Brechbiel, Martin W. ; King, Dylan ; Spirkoska, Violeta ; Allan, Laura C. ; Ramsland, Paul A. ; Scott, Andrew M. / Cross-species analysis of Fc engineered anti-Lewis-Y human IgG1 variants in human neonatal receptor transgenic mice reveal importance of S254 and Y436 in binding human neonatal Fc receptor. In: mAbs. 2016 ; Vol. 8, No. 4. pp. 775-786.
@article{6a4208db649d4c6bbc85e6d7a59e24a6,
title = "Cross-species analysis of Fc engineered anti-Lewis-Y human IgG1 variants in human neonatal receptor transgenic mice reveal importance of S254 and Y436 in binding human neonatal Fc receptor",
abstract = "IgG has a long half-life through engagement of its Fc region with the neonatal Fc receptor (FcRn). The FcRn binding site on IgG1 has been shown to contain I253 and H310 in the CH2 domain and H435 in the CH3 domain. Altering the half-life of IgG has been pursued with the aim to prolong or reduce the half-life of therapeutic IgGs. More recent studies have shown that IgGs bind differently to mouse and human FcRn. In this study we characterize a set of hu3S193 IgG1 variants with mutations in the FcRn binding site. A double mutation in the binding site is necessary to abrogate binding to murine FcRn, whereas a single mutation in the FcRn binding site is sufficient to no longer detect binding to human FcRn and create hu3S193 IgG1 variants with a half-life similar to previously studied hu3S193 F(ab’)2 (t1/2β, I253A, 12.23 h; H310A, 12.94; H435A, 12.57; F(ab’)2, 12.6 h). Alanine substitutions in S254 in the CH2 domain and Y436 in the CH3 domain showed reduced binding in vitro to human FcRn and reduced elimination half-lives in huFcRn transgenic mice (t1/2β, S254A, 37.43 h; Y436A, 39.53 h; wild-type, 83.15 h). These variants had minimal effect on half-life in BALB/c nu/nu mice (t1/2β, S254A, 119.9 h; Y436A, 162.1 h; wild-type, 163.1 h). These results provide insight into the interaction of human Fc by human FcRn, and are important for antibody-based therapeutics with optimal pharmacokinetics for payload strategies used in the clinic.",
keywords = "Antibody engineering, Fc receptors, Molecular biology, Neonatal Fc receptor, Transgenic mice",
author = "Burvenich, {Ingrid J.G.} and William Farrugia and Lee, {Fook T.} and Bruno Catimel and Zhanqi Liu and Dahna Makris and Diana Cao and O'Keefe, {Graeme J.} and Brechbiel, {Martin W.} and Dylan King and Violeta Spirkoska and Allan, {Laura C.} and Ramsland, {Paul A.} and Scott, {Andrew M.}",
year = "2016",
month = "5",
day = "18",
doi = "10.1080/19420862.2016.1156285",
language = "English",
volume = "8",
pages = "775--786",
journal = "mAbs",
issn = "1942-0862",
publisher = "Taylor & Francis",
number = "4",

}

Burvenich, IJG, Farrugia, W, Lee, FT, Catimel, B, Liu, Z, Makris, D, Cao, D, O'Keefe, GJ, Brechbiel, MW, King, D, Spirkoska, V, Allan, LC, Ramsland, PA & Scott, AM 2016, 'Cross-species analysis of Fc engineered anti-Lewis-Y human IgG1 variants in human neonatal receptor transgenic mice reveal importance of S254 and Y436 in binding human neonatal Fc receptor', mAbs, vol. 8, no. 4, pp. 775-786. https://doi.org/10.1080/19420862.2016.1156285

Cross-species analysis of Fc engineered anti-Lewis-Y human IgG1 variants in human neonatal receptor transgenic mice reveal importance of S254 and Y436 in binding human neonatal Fc receptor. / Burvenich, Ingrid J.G.; Farrugia, William; Lee, Fook T.; Catimel, Bruno; Liu, Zhanqi; Makris, Dahna; Cao, Diana; O'Keefe, Graeme J.; Brechbiel, Martin W.; King, Dylan; Spirkoska, Violeta; Allan, Laura C.; Ramsland, Paul A.; Scott, Andrew M.

In: mAbs, Vol. 8, No. 4, 18.05.2016, p. 775-786.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Cross-species analysis of Fc engineered anti-Lewis-Y human IgG1 variants in human neonatal receptor transgenic mice reveal importance of S254 and Y436 in binding human neonatal Fc receptor

AU - Burvenich, Ingrid J.G.

AU - Farrugia, William

AU - Lee, Fook T.

AU - Catimel, Bruno

AU - Liu, Zhanqi

AU - Makris, Dahna

AU - Cao, Diana

AU - O'Keefe, Graeme J.

AU - Brechbiel, Martin W.

AU - King, Dylan

AU - Spirkoska, Violeta

AU - Allan, Laura C.

AU - Ramsland, Paul A.

AU - Scott, Andrew M.

PY - 2016/5/18

Y1 - 2016/5/18

N2 - IgG has a long half-life through engagement of its Fc region with the neonatal Fc receptor (FcRn). The FcRn binding site on IgG1 has been shown to contain I253 and H310 in the CH2 domain and H435 in the CH3 domain. Altering the half-life of IgG has been pursued with the aim to prolong or reduce the half-life of therapeutic IgGs. More recent studies have shown that IgGs bind differently to mouse and human FcRn. In this study we characterize a set of hu3S193 IgG1 variants with mutations in the FcRn binding site. A double mutation in the binding site is necessary to abrogate binding to murine FcRn, whereas a single mutation in the FcRn binding site is sufficient to no longer detect binding to human FcRn and create hu3S193 IgG1 variants with a half-life similar to previously studied hu3S193 F(ab’)2 (t1/2β, I253A, 12.23 h; H310A, 12.94; H435A, 12.57; F(ab’)2, 12.6 h). Alanine substitutions in S254 in the CH2 domain and Y436 in the CH3 domain showed reduced binding in vitro to human FcRn and reduced elimination half-lives in huFcRn transgenic mice (t1/2β, S254A, 37.43 h; Y436A, 39.53 h; wild-type, 83.15 h). These variants had minimal effect on half-life in BALB/c nu/nu mice (t1/2β, S254A, 119.9 h; Y436A, 162.1 h; wild-type, 163.1 h). These results provide insight into the interaction of human Fc by human FcRn, and are important for antibody-based therapeutics with optimal pharmacokinetics for payload strategies used in the clinic.

AB - IgG has a long half-life through engagement of its Fc region with the neonatal Fc receptor (FcRn). The FcRn binding site on IgG1 has been shown to contain I253 and H310 in the CH2 domain and H435 in the CH3 domain. Altering the half-life of IgG has been pursued with the aim to prolong or reduce the half-life of therapeutic IgGs. More recent studies have shown that IgGs bind differently to mouse and human FcRn. In this study we characterize a set of hu3S193 IgG1 variants with mutations in the FcRn binding site. A double mutation in the binding site is necessary to abrogate binding to murine FcRn, whereas a single mutation in the FcRn binding site is sufficient to no longer detect binding to human FcRn and create hu3S193 IgG1 variants with a half-life similar to previously studied hu3S193 F(ab’)2 (t1/2β, I253A, 12.23 h; H310A, 12.94; H435A, 12.57; F(ab’)2, 12.6 h). Alanine substitutions in S254 in the CH2 domain and Y436 in the CH3 domain showed reduced binding in vitro to human FcRn and reduced elimination half-lives in huFcRn transgenic mice (t1/2β, S254A, 37.43 h; Y436A, 39.53 h; wild-type, 83.15 h). These variants had minimal effect on half-life in BALB/c nu/nu mice (t1/2β, S254A, 119.9 h; Y436A, 162.1 h; wild-type, 163.1 h). These results provide insight into the interaction of human Fc by human FcRn, and are important for antibody-based therapeutics with optimal pharmacokinetics for payload strategies used in the clinic.

KW - Antibody engineering

KW - Fc receptors

KW - Molecular biology

KW - Neonatal Fc receptor

KW - Transgenic mice

UR - http://www.scopus.com/inward/record.url?scp=84962052882&partnerID=8YFLogxK

U2 - 10.1080/19420862.2016.1156285

DO - 10.1080/19420862.2016.1156285

M3 - Article

VL - 8

SP - 775

EP - 786

JO - mAbs

JF - mAbs

SN - 1942-0862

IS - 4

ER -