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

• Published in: ImmunoTargets and therapy Vol. 9; pp. 299 - 316
• Main Authors: 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
• Format: Journal Article
• Language: English
• Published: New Zealand Dove Medical Press Limited 2020; Taylor & Francis Ltd; Dove Press; Dove; Dove Medical Press
• Subjects: 60 APPLIED LIFE SCIENCES; Agglutination; Antibiotics; Antibodies; Antigens; Bacteria; Cells (Biology); Drug resistance; Enzyme-linked immunosorbent assay; Epithelial cells; Flow cytometry; Health aspects; immunoantibiotic; immunodiagnostic; Immunoglobulin G; Immunoreactivity; immunotherapy; lateral flow assay; LFA; Mammalian cells; Mass spectrometry; Mass spectroscopy; Microbial drug resistance; Monoclonal antibodies; Multidrug resistance; Mutation; National security; Original Research; Phages; Plague; Proteins; Public health; radioimmunotherapy; radiolabeling; RIT; Scientific equipment industry; Therapeutic applications; Therapeutics; Virulence; Yersinia pestis; United States; Massachusetts; United States > US; radioimmunotherapy; lateral flow assay; LFA; immunotherapy; immunodiagnostic; radiolabeling; RIT; immunoantibiotic
• ISSN: 2253-1556; 2253-1556
• DOI: 10.2147/ITT.S267077

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 as a potential biothreat agent. Therefore, high-quality antibodies designed for accurate and sensitive 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 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 , and retained immunoreactivity upon chemical derivatization. Most importantly, these antibodies detected <1,000 cells in sandwich ELISA, did not harm respiratory epithelial cells, induced 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.