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Title: S tructural basis of selectivity and neutralizing activity of a TGFα/epiregulin specific antibody

Abstract

Abstract Recent studies have implicated a role of the epidermal growth factor receptor (EGFR) pathway in kidney disease. Skin toxicity associated with therapeutics which completely block the EGFR pathway precludes their use in chronic dosing. Therefore, we developed antibodies which specifically neutralize the EGFR ligands TGFα (transforming growth factor‐alpha) and epiregulin but not EGF (epidermal growth factor), amphiregulin, betacellulin, HB‐EGF (heparin‐binding epidermal growth factor), or epigen. The epitope of one such neutralizing antibody, LY3016859, was characterized in detail to elucidate the structural basis for ligand specificity. Here we report a crystal structure of the LY3016859 Fab fragment in complex with soluble human TGFα. Our data demonstrate a conformational epitope located primarily within the C‐terminal subdomain of the ligand. In addition, point mutagenesis experiments were used to highlight specific amino acids which are critical for both antigen binding and neutralization, most notably Ala 41 , Glu 44 , and His 45 . These results illustrate the structural basis for the ligand specificity/selectivity of LY3016859 and could also provide insight into further engineering to alter specificity and/or affinity of LY3016859.

Authors:
 [1];  [2];  [2];  [1];  [1]
  1. Biotechnology Discovery Research, Lilly Research Laboratories, Eli Lilly and Company Indianapolis Indiana 46285
  2. Eli Lilly Biotechnology Center San Diego California 92121
Publication Date:
Sponsoring Org.:
USDOE
OSTI Identifier:
1401704
Grant/Contract Number:  
DE‐AC02‐06CH11357
Resource Type:
Publisher's Accepted Manuscript
Journal Name:
Protein Science
Additional Journal Information:
Journal Name: Protein Science Journal Volume: 25 Journal Issue: 11; Journal ID: ISSN 0961-8368
Publisher:
Wiley Blackwell (John Wiley & Sons)
Country of Publication:
United Kingdom
Language:
English

Citation Formats

Boyles, Jeffrey S., Atwell, Shane, Druzina, Zhanna, Heuer, Josef G., and Witcher, Derrick R. S tructural basis of selectivity and neutralizing activity of a TGFα/epiregulin specific antibody. United Kingdom: N. p., 2016. Web. doi:10.1002/pro.3023.
Boyles, Jeffrey S., Atwell, Shane, Druzina, Zhanna, Heuer, Josef G., & Witcher, Derrick R. S tructural basis of selectivity and neutralizing activity of a TGFα/epiregulin specific antibody. United Kingdom. https://doi.org/10.1002/pro.3023
Boyles, Jeffrey S., Atwell, Shane, Druzina, Zhanna, Heuer, Josef G., and Witcher, Derrick R. Fri . "S tructural basis of selectivity and neutralizing activity of a TGFα/epiregulin specific antibody". United Kingdom. https://doi.org/10.1002/pro.3023.
@article{osti_1401704,
title = {S tructural basis of selectivity and neutralizing activity of a TGFα/epiregulin specific antibody},
author = {Boyles, Jeffrey S. and Atwell, Shane and Druzina, Zhanna and Heuer, Josef G. and Witcher, Derrick R.},
abstractNote = {Abstract Recent studies have implicated a role of the epidermal growth factor receptor (EGFR) pathway in kidney disease. Skin toxicity associated with therapeutics which completely block the EGFR pathway precludes their use in chronic dosing. Therefore, we developed antibodies which specifically neutralize the EGFR ligands TGFα (transforming growth factor‐alpha) and epiregulin but not EGF (epidermal growth factor), amphiregulin, betacellulin, HB‐EGF (heparin‐binding epidermal growth factor), or epigen. The epitope of one such neutralizing antibody, LY3016859, was characterized in detail to elucidate the structural basis for ligand specificity. Here we report a crystal structure of the LY3016859 Fab fragment in complex with soluble human TGFα. Our data demonstrate a conformational epitope located primarily within the C‐terminal subdomain of the ligand. In addition, point mutagenesis experiments were used to highlight specific amino acids which are critical for both antigen binding and neutralization, most notably Ala 41 , Glu 44 , and His 45 . These results illustrate the structural basis for the ligand specificity/selectivity of LY3016859 and could also provide insight into further engineering to alter specificity and/or affinity of LY3016859.},
doi = {10.1002/pro.3023},
journal = {Protein Science},
number = 11,
volume = 25,
place = {United Kingdom},
year = {Fri Sep 09 00:00:00 EDT 2016},
month = {Fri Sep 09 00:00:00 EDT 2016}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
https://doi.org/10.1002/pro.3023

Citation Metrics:
Cited by: 4 works
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