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Title: Development of Anti-Yersinia pestis Human Antibodies with Features Required for Diagnostic and Therapeutic Applications

Abstract

Yersinia pestis is a category A infective agent that causes bubonic, septicemic, and pneumonic plague. Notably, the acquisition of antimicrobial or multidrug resistance through natural or purposed means qualifies Y. pestis as a potential biothreat agent. Therefore, high-quality antibodies designed for accurate and sensitive Y. pestis diagnostics, and therapeutics potentiating or replacing traditional antibiotics are of utmost need for national security and public health preparedness. Here, we describe a set of human monoclonal immunoglobulins (IgG1s) targeting Y. pestis fraction 1 (F1) antigen, previously derived from in vitro evolution of a phage-display library of single-chain antibodies (scFv). We extensively characterized these antibodies and their effect on bacterial and mammalian cells via: ELISA, flow cytometry, mass spectrometry, spectroscopy, and various metabolic assays. Two of our anti-F1 IgG (αF1Ig 2 and αF1Ig 8) stood out for high production yield, specificity, and stability. These two antibodies were additionally attractive in that they displayed picomolar affinity, did not compete when binding Y. pestis, and retained immunoreactivity upon chemical derivatization. Most importantly, these antibodies detected < 1,000 Y. pestis cells in sandwich ELISA, did not harm respiratory epithelial cells, induced Y. pestis agglutination at low concentration (350 nM), and caused apparent reduction in cell growth whenmore » radiolabeled at a nonagglutinating concentration (34 nM). These antibodies are amenable to the development of accurate and sensitive diagnostics and immuno/radioimmunotherapeutics.« less

Authors:
ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [1];  [1]; ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [1];  [1];  [1]; ORCiD logo [2]; ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [1]
  1. Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
  2. Specifica Inc., Santa Fe, NM (United States)
Publication Date:
Research Org.:
Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
Sponsoring Org.:
USDOE Laboratory Directed Research and Development (LDRD) Program; USDOE Office of Science (SC), Nuclear Physics (NP)
OSTI Identifier:
1755889
Report Number(s):
LA-UR-20-30360
Journal ID: ISSN 2253-1556
Grant/Contract Number:  
89233218CNA000001; 20180005DR
Resource Type:
Accepted Manuscript
Journal Name:
ImmunoTargets and Therapy
Additional Journal Information:
Journal Volume: 9; Journal ID: ISSN 2253-1556
Publisher:
Dove Press
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; immunodiagnostic; radioimmunotherapy; RIT; immunotherapy; radiolabeling; immunoantibiotic; lateral flow assay; LFA

Citation Formats

Lillo, Antonietta M., Velappan, Nileena, Kelliher, Julia M., Watts, Austin J., Merriman, Samuel P., Vuyisich, Grace, Lilley, Laura M., Coombs, Kent E., Mastren, Tara, Teshima, Munehiro, Stein, Benjamin W., Wagner, Gregory L., Iyer, Srinivas, Bradbury, Andrew R. M., Harris, Jennifer Foster, Dichosa, Armand E., and Kozimor, Stosh A.. Development of Anti-Yersinia pestis Human Antibodies with Features Required for Diagnostic and Therapeutic Applications. United States: N. p., 2020. Web. https://doi.org/10.2147/itt.s267077.
Lillo, Antonietta M., Velappan, Nileena, Kelliher, Julia M., Watts, Austin J., Merriman, Samuel P., Vuyisich, Grace, Lilley, Laura M., Coombs, Kent E., Mastren, Tara, Teshima, Munehiro, Stein, Benjamin W., Wagner, Gregory L., Iyer, Srinivas, Bradbury, Andrew R. M., Harris, Jennifer Foster, Dichosa, Armand E., & Kozimor, Stosh A.. Development of Anti-Yersinia pestis Human Antibodies with Features Required for Diagnostic and Therapeutic Applications. United States. https://doi.org/10.2147/itt.s267077
Lillo, Antonietta M., Velappan, Nileena, Kelliher, Julia M., Watts, Austin J., Merriman, Samuel P., Vuyisich, Grace, Lilley, Laura M., Coombs, Kent E., Mastren, Tara, Teshima, Munehiro, Stein, Benjamin W., Wagner, Gregory L., Iyer, Srinivas, Bradbury, Andrew R. M., Harris, Jennifer Foster, Dichosa, Armand E., and Kozimor, Stosh A.. Fri . "Development of Anti-Yersinia pestis Human Antibodies with Features Required for Diagnostic and Therapeutic Applications". United States. https://doi.org/10.2147/itt.s267077. https://www.osti.gov/servlets/purl/1755889.
@article{osti_1755889,
title = {Development of Anti-Yersinia pestis Human Antibodies with Features Required for Diagnostic and Therapeutic Applications},
author = {Lillo, Antonietta M. and Velappan, Nileena and Kelliher, Julia M. and Watts, Austin J. and Merriman, Samuel P. and Vuyisich, Grace and Lilley, Laura M. and Coombs, Kent E. and Mastren, Tara and Teshima, Munehiro and Stein, Benjamin W. and Wagner, Gregory L. and Iyer, Srinivas and Bradbury, Andrew R. M. and Harris, Jennifer Foster and Dichosa, Armand E. and Kozimor, Stosh A.},
abstractNote = {Yersinia pestis is a category A infective agent that causes bubonic, septicemic, and pneumonic plague. Notably, the acquisition of antimicrobial or multidrug resistance through natural or purposed means qualifies Y. pestis as a potential biothreat agent. Therefore, high-quality antibodies designed for accurate and sensitive Y. pestis diagnostics, and therapeutics potentiating or replacing traditional antibiotics are of utmost need for national security and public health preparedness. Here, we describe a set of human monoclonal immunoglobulins (IgG1s) targeting Y. pestis fraction 1 (F1) antigen, previously derived from in vitro evolution of a phage-display library of single-chain antibodies (scFv). We extensively characterized these antibodies and their effect on bacterial and mammalian cells via: ELISA, flow cytometry, mass spectrometry, spectroscopy, and various metabolic assays. Two of our anti-F1 IgG (αF1Ig 2 and αF1Ig 8) stood out for high production yield, specificity, and stability. These two antibodies were additionally attractive in that they displayed picomolar affinity, did not compete when binding Y. pestis, and retained immunoreactivity upon chemical derivatization. Most importantly, these antibodies detected < 1,000 Y. pestis cells in sandwich ELISA, did not harm respiratory epithelial cells, induced Y. pestis agglutination at low concentration (350 nM), and caused apparent reduction in cell growth when radiolabeled at a nonagglutinating concentration (34 nM). These antibodies are amenable to the development of accurate and sensitive diagnostics and immuno/radioimmunotherapeutics.},
doi = {10.2147/itt.s267077},
journal = {ImmunoTargets and Therapy},
number = ,
volume = 9,
place = {United States},
year = {2020},
month = {11}
}

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