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Title: De Novo Sequencing and Resurrection of a Human Astrovirus-Neutralizing Antibody

Abstract

Monoclonal antibody (mAb) therapeutics targeting cancer, autoimmune diseases, inflammatory diseases, and infectious diseases are growing exponentially. Although numerous panels of mAbs targeting infectious disease agents have been developed, their progression into clinically useful mAbs is often hindered by the lack of sequence information and/or loss of hybridoma cells that produce them. Here we combine the power of crystallography and mass spectrometry to determine the amino acid sequence and glycosylation modification of the Fab fragment of a potent human astrovirus-neutralizing mAb. We used this information to engineer a recombinant antibody single-chain variable fragment that has the same specificity as the parent monoclonal antibody to bind to the astrovirus capsid protein. Furthermore, this antibody can now potentially be developed as a therapeutic and diagnostic agent.

Authors:
 [1];  [2];  [3];  [2];  [4];  [1]
+ Show Author Affiliations
  1. Department of Biomolecular Engineering, University of California Santa Cruz, 1156 High Street, Santa Cruz, California 95064, United States
  2. Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, New York, New York 10016, United States
  3. Protein Metrics, San Carlos, California 94070, United States
  4. Viral Gastroenteritis Unit, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Madrid, Spain
Publication Date:
Research Org.:
Univ. of California, Santa Cruz, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES)
OSTI Identifier:
1244771
Alternate Identifier(s):
OSTI ID: 1345043
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Published Article
Journal Name:
ACS Infectious Diseases
Additional Journal Information:
Journal Name: ACS Infectious Diseases Journal Volume: 2 Journal Issue: 5; Journal ID: ISSN 2373-8227
Publisher:
American Chemical Society
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; antibody; astrovirus; mass spectrometry; protein engineering; protein sequencing; X-ray crystallography

Citation Formats

Bogdanoff, Walter A., Morgenstern, David, Bern, Marshall, Ueberheide, Beatrix M., Sanchez-Fauquier, Alicia, and DuBois, Rebecca M. De Novo Sequencing and Resurrection of a Human Astrovirus-Neutralizing Antibody. United States: N. p., 2016. Web. doi:10.1021/acsinfecdis.6b00026.
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Bogdanoff, Walter A., Morgenstern, David, Bern, Marshall, Ueberheide, Beatrix M., Sanchez-Fauquier, Alicia, & DuBois, Rebecca M. De Novo Sequencing and Resurrection of a Human Astrovirus-Neutralizing Antibody. United States. https://doi.org/10.1021/acsinfecdis.6b00026
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Bogdanoff, Walter A., Morgenstern, David, Bern, Marshall, Ueberheide, Beatrix M., Sanchez-Fauquier, Alicia, and DuBois, Rebecca M. Thu . "De Novo Sequencing and Resurrection of a Human Astrovirus-Neutralizing Antibody". United States. https://doi.org/10.1021/acsinfecdis.6b00026.
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@article{osti_1244771,
title = {De Novo Sequencing and Resurrection of a Human Astrovirus-Neutralizing Antibody},
author = {Bogdanoff, Walter A. and Morgenstern, David and Bern, Marshall and Ueberheide, Beatrix M. and Sanchez-Fauquier, Alicia and DuBois, Rebecca M.},
abstractNote = {Monoclonal antibody (mAb) therapeutics targeting cancer, autoimmune diseases, inflammatory diseases, and infectious diseases are growing exponentially. Although numerous panels of mAbs targeting infectious disease agents have been developed, their progression into clinically useful mAbs is often hindered by the lack of sequence information and/or loss of hybridoma cells that produce them. Here we combine the power of crystallography and mass spectrometry to determine the amino acid sequence and glycosylation modification of the Fab fragment of a potent human astrovirus-neutralizing mAb. We used this information to engineer a recombinant antibody single-chain variable fragment that has the same specificity as the parent monoclonal antibody to bind to the astrovirus capsid protein. Furthermore, this antibody can now potentially be developed as a therapeutic and diagnostic agent.},
doi = {10.1021/acsinfecdis.6b00026},
journal = {ACS Infectious Diseases},
number = 5,
volume = 2,
place = {United States},
year = {Thu Mar 31 00:00:00 EDT 2016},
month = {Thu Mar 31 00:00:00 EDT 2016}
}
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Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
https://doi.org/10.1021/acsinfecdis.6b00026
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