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Title: Computational design of a specific heavy chain/κ light chain interface for expressing fully IgG bispecific antibodies

Abstract

Abstract The use of bispecific antibodies (BsAbs) to treat human diseases is on the rise. Increasingly complex and powerful therapeutic mechanisms made possible by BsAbs are spurring innovation of novel BsAb formats and methods for their production. The long‐lived in vivo pharmacokinetics, optimal biophysical properties and potential effector functions of natural IgG monoclonal (and monospecific) antibodies has resulted in a push to generate fully IgG BsAb formats with the same quaternary structure as monoclonal IgGs. The production of fully IgG BsAbs is challenging because of the highly heterogeneous pairing of heavy chains (HCs) and light chains (LCs) when produced in mammalian cells with two IgG HCs and two LCs. A solution to the HC heterodimerization aspect of IgG BsAb production was first discovered two decades ago; however, addressing the LC mispairing issue has remained intractable until recently. Here, we use computational and rational engineering to develop novel designs to the HC/LC pairing issue, and particularly for κ LCs. Crystal structures of these designs highlight the interactions that provide HC/LC specificity. We produce and characterize multiple fully IgG BsAbs using these novel designs. We demonstrate the importance of specificity engineering in both the variable and constant domains to achieve robust HC/LCmore » specificity within all the BsAbs. These solutions facilitate the production of fully IgG BsAbs for clinical use.« less

Authors:
 [1];  [2];  [1];  [1];  [1];  [1];  [1];  [1];  [2];  [1];  [1];  [1];  [3];  [1]
+ Show Author Affiliations
  1. Eli Lilly Biotechnology Center, 10300 Campus Point Drive San Diego California 92121
  2. Department of Biochemistry and Biophysics University of North Carolina at Chapel Hill Chapel Hill North Carolina
  3. Department of Biochemistry and Biophysics University of North Carolina at Chapel Hill Chapel Hill North Carolina, Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill Chapel Hill North Carolina
Publication Date:
Sponsoring Org.:
USDOE
OSTI Identifier:
1393306
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: 26 Journal Issue: 10; Journal ID: ISSN 0961-8368
Publisher:
Wiley Blackwell (John Wiley & Sons)
Country of Publication:
United Kingdom
Language:
English

Citation Formats

Froning, K. J., Leaver‐Fay, A., Wu, X., Phan, S., Gao, L., Huang, F., Pustilnik, A., Bacica, M., Houlihan, K., Chai, Q., Fitchett, J. R., Hendle, J., Kuhlman, B., and Demarest, S. J. Computational design of a specific heavy chain/κ light chain interface for expressing fully IgG bispecific antibodies. United Kingdom: N. p., 2017. Web. doi:10.1002/pro.3240.
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Froning, K. J., Leaver‐Fay, A., Wu, X., Phan, S., Gao, L., Huang, F., Pustilnik, A., Bacica, M., Houlihan, K., Chai, Q., Fitchett, J. R., Hendle, J., Kuhlman, B., & Demarest, S. J. Computational design of a specific heavy chain/κ light chain interface for expressing fully IgG bispecific antibodies. United Kingdom. https://doi.org/10.1002/pro.3240
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Froning, K. J., Leaver‐Fay, A., Wu, X., Phan, S., Gao, L., Huang, F., Pustilnik, A., Bacica, M., Houlihan, K., Chai, Q., Fitchett, J. R., Hendle, J., Kuhlman, B., and Demarest, S. J. Mon . "Computational design of a specific heavy chain/κ light chain interface for expressing fully IgG bispecific antibodies". United Kingdom. https://doi.org/10.1002/pro.3240.
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@article{osti_1393306,
title = {Computational design of a specific heavy chain/κ light chain interface for expressing fully IgG bispecific antibodies},
author = {Froning, K. J. and Leaver‐Fay, A. and Wu, X. and Phan, S. and Gao, L. and Huang, F. and Pustilnik, A. and Bacica, M. and Houlihan, K. and Chai, Q. and Fitchett, J. R. and Hendle, J. and Kuhlman, B. and Demarest, S. J.},
abstractNote = {Abstract The use of bispecific antibodies (BsAbs) to treat human diseases is on the rise. Increasingly complex and powerful therapeutic mechanisms made possible by BsAbs are spurring innovation of novel BsAb formats and methods for their production. The long‐lived in vivo pharmacokinetics, optimal biophysical properties and potential effector functions of natural IgG monoclonal (and monospecific) antibodies has resulted in a push to generate fully IgG BsAb formats with the same quaternary structure as monoclonal IgGs. The production of fully IgG BsAbs is challenging because of the highly heterogeneous pairing of heavy chains (HCs) and light chains (LCs) when produced in mammalian cells with two IgG HCs and two LCs. A solution to the HC heterodimerization aspect of IgG BsAb production was first discovered two decades ago; however, addressing the LC mispairing issue has remained intractable until recently. Here, we use computational and rational engineering to develop novel designs to the HC/LC pairing issue, and particularly for κ LCs. Crystal structures of these designs highlight the interactions that provide HC/LC specificity. We produce and characterize multiple fully IgG BsAbs using these novel designs. We demonstrate the importance of specificity engineering in both the variable and constant domains to achieve robust HC/LC specificity within all the BsAbs. These solutions facilitate the production of fully IgG BsAbs for clinical use.},
doi = {10.1002/pro.3240},
journal = {Protein Science},
number = 10,
volume = 26,
place = {United Kingdom},
year = {Mon Jul 31 00:00:00 EDT 2017},
month = {Mon Jul 31 00:00:00 EDT 2017}
}
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https://doi.org/10.1002/pro.3240
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