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Title: Optimization of the Solubility of HIV-1-Neutralizing Antibody 10E8 through Somatic Variation and Structure-Based Design

Abstract

Extraordinary antibodies capable of near pan-neutralization of HIV-1 have been identified. One of the broadest is antibody 10E8, which recognizes the membrane-proximal external region (MPER) of the HIV-1 envelope and neutralizes >95% of circulating HIV-1 strains. If delivered passively, 10E8 might serve to prevent or treat HIV-1 infection. Antibody 10E8, however, is markedly less soluble than other antibodies. Here, we describe the use of both structural biology and somatic variation to develop optimized versions of 10E8 with increased solubility. From the structure of 10E8, we identified a prominent hydrophobic patch; reversion of four hydrophobic residues in this patch to their hydrophilic germ line counterparts resulted in an ~10-fold decrease in turbidity. We also used somatic variants of 10E8, identified previously by next-generation sequencing, to optimize heavy and light chains; this process yielded several improved variants. Of these, variant 10E8v4 with 26 changes versus the parent 10E8 was the most soluble, with a paratope we showed crystallographically to be virtually identical to that of 10E8, a potency on a panel of 200 HIV-1 isolates also similar to that of 10E8, and a half-life in rhesus macaques of ~10 days. An anomaly in 10E8v4 size exclusion chromatography that appeared to be relatedmore » to conformational isomerization was resolved by engineering an interchain disulfide. Furthermore, by combining a structure-based approach with natural variation in potency and solubility from the 10E8 lineage, we successfully created variants of 10E8 which retained the potency and extraordinary neutralization breadth of the parent 10E8 but with substantially increased solubility.« less

Authors:
 [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1] more »;  [1];  [1];  [1];  [1];  [2];  [2];  [2];  [2];  [2];  [2];  [3];  [3];  [3];  [1];  [1];  [1];  [1];  [1];  [4] « less
  1. National Inst. of Health (NIH), Bethesda, MD (United States)
  2. Duke Univ. School of Medicine, Durham, NC (United States); Duke Univ., Durham, NC (United States)
  3. Rockefeller Univ., New York, NY (United States)
  4. Univ. of Utah, Salt Lake City, UT (United States)
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
OSTI Identifier:
1314247
Grant/Contract Number:  
W-31-109-Eng-38
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Journal of Virology
Additional Journal Information:
Journal Volume: 90; Journal Issue: 13; Journal ID: ISSN 0022-538X
Publisher:
American Society for Microbiology
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Kwon, Young D., Georgiev, Ivelin S., Ofek, Gilad, Zhang, Baoshan, Asokan, Mangaiarkarasi, Bailer, Robert T., Bao, Amy, Caruso, William, Chen, Xuejun, Choe, Misook, Druz, Aliaksandr, Ko, Sung-Youl, Louder, Mark K., McKee, Krisha, O'Dell, Sijy, Pegu, Amarendra, Rudicell, Rebecca S., Shi, Wei, Wang, Keyun, Yang, Yongping, Alger, Mandy, Bender, Michael F., Carlton, Kevin, Cooper, Jonathan W., Blinn, Julie, Eudailey, Joshua, Lloyd, Krissey, Parks, Robert, Alam, S. Munir, Haynes, Barton F., Padte, Neal N., Yu, Jian, Ho, David D., Huang, Jinghe, Connors, Mark, Schwartz, Richard M., Mascola, John R., Kwong, Peter D., and Sundquist, W. I. Optimization of the Solubility of HIV-1-Neutralizing Antibody 10E8 through Somatic Variation and Structure-Based Design. United States: N. p., 2016. Web. doi:10.1128/JVI.03246-15.
Kwon, Young D., Georgiev, Ivelin S., Ofek, Gilad, Zhang, Baoshan, Asokan, Mangaiarkarasi, Bailer, Robert T., Bao, Amy, Caruso, William, Chen, Xuejun, Choe, Misook, Druz, Aliaksandr, Ko, Sung-Youl, Louder, Mark K., McKee, Krisha, O'Dell, Sijy, Pegu, Amarendra, Rudicell, Rebecca S., Shi, Wei, Wang, Keyun, Yang, Yongping, Alger, Mandy, Bender, Michael F., Carlton, Kevin, Cooper, Jonathan W., Blinn, Julie, Eudailey, Joshua, Lloyd, Krissey, Parks, Robert, Alam, S. Munir, Haynes, Barton F., Padte, Neal N., Yu, Jian, Ho, David D., Huang, Jinghe, Connors, Mark, Schwartz, Richard M., Mascola, John R., Kwong, Peter D., & Sundquist, W. I. Optimization of the Solubility of HIV-1-Neutralizing Antibody 10E8 through Somatic Variation and Structure-Based Design. United States. doi:10.1128/JVI.03246-15.
Kwon, Young D., Georgiev, Ivelin S., Ofek, Gilad, Zhang, Baoshan, Asokan, Mangaiarkarasi, Bailer, Robert T., Bao, Amy, Caruso, William, Chen, Xuejun, Choe, Misook, Druz, Aliaksandr, Ko, Sung-Youl, Louder, Mark K., McKee, Krisha, O'Dell, Sijy, Pegu, Amarendra, Rudicell, Rebecca S., Shi, Wei, Wang, Keyun, Yang, Yongping, Alger, Mandy, Bender, Michael F., Carlton, Kevin, Cooper, Jonathan W., Blinn, Julie, Eudailey, Joshua, Lloyd, Krissey, Parks, Robert, Alam, S. Munir, Haynes, Barton F., Padte, Neal N., Yu, Jian, Ho, David D., Huang, Jinghe, Connors, Mark, Schwartz, Richard M., Mascola, John R., Kwong, Peter D., and Sundquist, W. I. Fri . "Optimization of the Solubility of HIV-1-Neutralizing Antibody 10E8 through Somatic Variation and Structure-Based Design". United States. doi:10.1128/JVI.03246-15. https://www.osti.gov/servlets/purl/1314247.
@article{osti_1314247,
title = {Optimization of the Solubility of HIV-1-Neutralizing Antibody 10E8 through Somatic Variation and Structure-Based Design},
author = {Kwon, Young D. and Georgiev, Ivelin S. and Ofek, Gilad and Zhang, Baoshan and Asokan, Mangaiarkarasi and Bailer, Robert T. and Bao, Amy and Caruso, William and Chen, Xuejun and Choe, Misook and Druz, Aliaksandr and Ko, Sung-Youl and Louder, Mark K. and McKee, Krisha and O'Dell, Sijy and Pegu, Amarendra and Rudicell, Rebecca S. and Shi, Wei and Wang, Keyun and Yang, Yongping and Alger, Mandy and Bender, Michael F. and Carlton, Kevin and Cooper, Jonathan W. and Blinn, Julie and Eudailey, Joshua and Lloyd, Krissey and Parks, Robert and Alam, S. Munir and Haynes, Barton F. and Padte, Neal N. and Yu, Jian and Ho, David D. and Huang, Jinghe and Connors, Mark and Schwartz, Richard M. and Mascola, John R. and Kwong, Peter D. and Sundquist, W. I.},
abstractNote = {Extraordinary antibodies capable of near pan-neutralization of HIV-1 have been identified. One of the broadest is antibody 10E8, which recognizes the membrane-proximal external region (MPER) of the HIV-1 envelope and neutralizes >95% of circulating HIV-1 strains. If delivered passively, 10E8 might serve to prevent or treat HIV-1 infection. Antibody 10E8, however, is markedly less soluble than other antibodies. Here, we describe the use of both structural biology and somatic variation to develop optimized versions of 10E8 with increased solubility. From the structure of 10E8, we identified a prominent hydrophobic patch; reversion of four hydrophobic residues in this patch to their hydrophilic germ line counterparts resulted in an ~10-fold decrease in turbidity. We also used somatic variants of 10E8, identified previously by next-generation sequencing, to optimize heavy and light chains; this process yielded several improved variants. Of these, variant 10E8v4 with 26 changes versus the parent 10E8 was the most soluble, with a paratope we showed crystallographically to be virtually identical to that of 10E8, a potency on a panel of 200 HIV-1 isolates also similar to that of 10E8, and a half-life in rhesus macaques of ~10 days. An anomaly in 10E8v4 size exclusion chromatography that appeared to be related to conformational isomerization was resolved by engineering an interchain disulfide. Furthermore, by combining a structure-based approach with natural variation in potency and solubility from the 10E8 lineage, we successfully created variants of 10E8 which retained the potency and extraordinary neutralization breadth of the parent 10E8 but with substantially increased solubility.},
doi = {10.1128/JVI.03246-15},
journal = {Journal of Virology},
issn = {0022-538X},
number = 13,
volume = 90,
place = {United States},
year = {2016},
month = {6}
}

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