{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,14]],"date-time":"2026-05-14T11:28:42Z","timestamp":1778758122462,"version":"3.51.4"},"reference-count":64,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2021,3,2]],"date-time":"2021-03-02T00:00:00Z","timestamp":1614643200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>Mouse monoclonal antibodies were raised against plague disease biomarkers: the bacterial capsular protein fraction 1 (F1) and the low-calcium response\u2014LcrV virulence factor (Vag). A novel tandem assay, employing BioLayer Interferometry (BLI), enabled the isolation of antibodies against four different epitopes on Vag. The tandem assay was carried out with hybridoma supernatants, circumventing the need for antibody purification. The BioLayer assay was further adopted for characterization of epitope-repetitive antigens, enabling the discovery of two unique epitopes on F1. The selected antibodies were purified and applied as \u201coligo-clonal\u201d reagents for the immuno-detection of both biomarkers. The developed Homogenous Time Resolved Fluorescence (HTRF) tests were short (10 min) and simple (no washing steps), allowing for detection of 10 ng\/mL F1 and 2.5 ng\/mL Vag. The tests were successfully applied for detection of disease biomarkers produced by various Y. pestis strains during growth in blood culture vials.<\/jats:p>","DOI":"10.3390\/pathogens10030285","type":"journal-article","created":{"date-parts":[[2021,3,2]],"date-time":"2021-03-02T10:36:37Z","timestamp":1614681397000},"page":"285","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Epitope Binning of Novel Monoclonal Anti F1 and Anti LcrV Antibodies and Their Application in a Simple, Short, HTRF Test for Clinical Plague Detection"],"prefix":"10.3390","volume":"10","author":[{"given":"Adva","family":"Mechaly","sequence":"first","affiliation":[{"name":"The Department of Infectious Diseases, Israel Institute for Biological Research, Ness-Ziona 7410001, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Einat B.","family":"Vitner","sequence":"additional","affiliation":[{"name":"The Department of Infectious Diseases, Israel Institute for Biological Research, Ness-Ziona 7410001, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yinon","family":"Levy","sequence":"additional","affiliation":[{"name":"The Department of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, Ness Ziona 7410001, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David","family":"Gur","sequence":"additional","affiliation":[{"name":"The Department of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, Ness Ziona 7410001, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Moria","family":"Barlev-Gross","sequence":"additional","affiliation":[{"name":"The Department of Infectious Diseases, Israel Institute for Biological Research, Ness-Ziona 7410001, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Assa","family":"Sittner","sequence":"additional","affiliation":[{"name":"The Department of Infectious Diseases, Israel Institute for Biological Research, Ness-Ziona 7410001, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michal","family":"Koren","sequence":"additional","affiliation":[{"name":"The Department of Infectious Diseases, Israel Institute for Biological Research, Ness-Ziona 7410001, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1902-8535","authenticated-orcid":false,"given":"Haim","family":"Levy","sequence":"additional","affiliation":[{"name":"The Department of Infectious Diseases, Israel Institute for Biological Research, Ness-Ziona 7410001, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5685-293X","authenticated-orcid":false,"given":"Emanuelle","family":"Mamroud","sequence":"additional","affiliation":[{"name":"The Department of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, Ness Ziona 7410001, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Morly","family":"Fisher","sequence":"additional","affiliation":[{"name":"The Department of Infectious Diseases, Israel Institute for Biological Research, Ness-Ziona 7410001, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,3,2]]},"reference":[{"key":"ref_1","first-page":"543","article-title":"Energy transfer efficacy as a function of donor-acceptor distance in a homogenous time resolved fluorescence energy transfer (htrf) immunoassay for protective antigen detection","volume":"27","author":"Cohen","year":"2012","journal-title":"Luminescence"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"5179","DOI":"10.1007\/s00216-016-9602-0","article-title":"An internal standard approach for homogeneous TR-FRET immunoassays facilitates the detection of bacteria, biomarkers, and toxins in complex matrices","volume":"408","author":"Cohen","year":"2016","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1002\/andp.19484370105","article-title":"Zwischenmolekulare Energiewanderung und Fluoreszenz","volume":"437","year":"1948","journal-title":"Annalen der Physik"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2330","DOI":"10.1128\/AEM.69.4.2330-2339.2003","article-title":"Detection of frequency resonance energy transfer pair on double-labeled microsphere and Bacillus anthracis spores by flow cytometry","volume":"69","author":"Zahavy","year":"2003","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1016\/j.jim.2012.05.010","article-title":"Label-free epitope binning assays of monoclonal antibodies enable the identification of antigen heterogeneity","volume":"382","author":"Abdiche","year":"2012","journal-title":"J. Immunol. Methods"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Abdiche, Y.N., Miles, A., Eckman, J., Foletti, D., Van Blarcom, T.J., Yeung, Y.A., Pons, J., and Rajpal, A. (2014). High-throughput epitope binning assays on label-free array-based biosensors can yield exquisite epitope discrimination that facilitates the selection of monoclonal antibodies with functional activity. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0092451"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"270","DOI":"10.4161\/mabs.23049","article-title":"High throughput solution-based measurement of antibody-antigen affinity and epitope binning","volume":"5","author":"Estep","year":"2013","journal-title":"MAbs"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1016\/j.jim.2004.02.010","article-title":"A novel method of Multiplexed Competitive Antibody Binning for the characterization of monoclonal antibodies","volume":"288","author":"Jia","year":"2004","journal-title":"J. Immunol. Methods"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"110","DOI":"10.4161\/19420862.2014.985919","article-title":"Characterization of monoclonal antibody\u2019s binding kinetics using oblique-incidence reflectivity difference approach","volume":"7","author":"Liu","year":"2015","journal-title":"MAbs"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1016\/j.jim.2010.12.021","article-title":"Epitope binning of murine monoclonal antibodies by a multiplexed pairing assay","volume":"365","author":"Miller","year":"2011","journal-title":"J. Immunol. Methods"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1931","DOI":"10.1016\/j.molimm.2010.02.020","article-title":"Characterization of a panel of anti-tetanus toxin single-chain Fvs reveals cooperative binding","volume":"47","author":"Scott","year":"2010","journal-title":"Mol. Immunol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1128\/CMR.10.1.35","article-title":"Yersinia pestis\u2014Etiological agent of plague","volume":"10","author":"Perry","year":"1997","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2281","DOI":"10.1001\/jama.283.17.2281","article-title":"Plague as a biological weapon: Medical and public health management. Working Group on Civilian Biodefense","volume":"283","author":"Inglesby","year":"2000","journal-title":"JAMA"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1016\/S1438-4221(00)80126-5","article-title":"Early diagnosis of bubonic plague using F1 antigen capture ELISA assay and rapid immunogold dipstick","volume":"290","author":"Chanteau","year":"2000","journal-title":"Int. J. Med. Microbiol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1016\/j.femsim.2004.02.005","article-title":"Evaluation of a standardized F1 capsular antigen capture ELISA test kit for the rapid diagnosis of plague","volume":"41","author":"Splettstoesser","year":"2004","journal-title":"FEMS Immunol. Med. Microbiol."},{"key":"ref_16","first-page":"745","article-title":"Application of enzyme immunoassays for the confirmation of clinically suspect plague in Namibia","volume":"64","author":"Williams","year":"1986","journal-title":"Bull. WHO"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Rajerison, M., Melocco, M., Andrianaivoarimanana, V., Rahajandraibe, S., Rakotoarimanana, F., Spiegel, A., Ratsitorahina, M., and Baril, L. (2020). Performance of plague rapid diagnostic test compared to bacteriology: A retrospective analysis of the data collected in Madagascar. BMC Infect. Dis., 20.","DOI":"10.1186\/s12879-020-4812-7"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1111\/j.1574-695X.2010.00687.x","article-title":"The search for early markers of plague: Evidence for accumulation of soluble Yersinia pestis LcrV in bubonic and pneumonic mouse models of disease","volume":"59","author":"Flashner","year":"2010","journal-title":"FEMS Immunol. Med. Microbiol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"434","DOI":"10.1128\/CMR.17.2.434-464.2004","article-title":"Intraspecific diversity of Yersinia pestis","volume":"17","author":"Anisimov","year":"2004","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_20","first-page":"156","article-title":"Pathology of experimental pneumonic plague produced by F1-positive and F1-negative Yersinia pestis in african green monkeys","volume":"120","author":"Davies","year":"1996","journal-title":"Arch. Pathol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"262","DOI":"10.2105\/AJPH.68.3.262","article-title":"Atypical plague bacilli isolated from rodents, fleas, and man","volume":"68","author":"Williams","year":"1978","journal-title":"Am. J. Public Health"},{"key":"ref_22","first-page":"408","article-title":"An unusual strain of Pasteurella pestis isolated from a fatal human case of plague","volume":"23","author":"Winter","year":"1960","journal-title":"B World Health Organ."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"e100","DOI":"10.14440\/jbm.2018.257","article-title":"Improved production of monoclonal antibodies against the LcrV antigen of Yersinia pestis using FACS-aided hybridoma selection","volume":"5","author":"Sittner","year":"2018","journal-title":"J. Biol. Meth."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"498","DOI":"10.1177\/1087057114560123","article-title":"High-throughput kinetic screening of hybridomas to identify high-affinity antibodies using bio-layer interferometry","volume":"20","author":"Lad","year":"2015","journal-title":"J. Biomol. Screen"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"208","DOI":"10.1016\/j.ab.2013.07.025","article-title":"Off-rate screening for selection of high-affinity anti-drug antibodies","volume":"441","author":"Ylera","year":"2013","journal-title":"Anal. Biochem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2180","DOI":"10.1128\/iai.64.6.2180-2187.1996","article-title":"Fraction 1 capsular antigen (F1) purification from Yersinia pestis CO92 and from an Escherichia coli recombinant strain and efficacy against lethal plague challenge","volume":"64","author":"Andrews","year":"1996","journal-title":"Infect. Immun."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"587","DOI":"10.1016\/S0092-8674(03)00351-9","article-title":"Structure and biogenesis of the capsular F1 antigen from Yersinia pestis: Preserved folding energy drives fiber formation","volume":"113","author":"Zavialov","year":"2003","journal-title":"Cell"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"795","DOI":"10.1007\/s10895-014-1354-7","article-title":"Rapid homogenous time-resolved fluorescence (HTRF) immunoassay for anthrax detection","volume":"24","author":"Cohen","year":"2014","journal-title":"J. Fluoresc."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"524","DOI":"10.1134\/S0003683819040082","article-title":"Characteristics of the Chromatographic Cleaning and Protectiveness of the LcrV Isoform of Yersinia pestis","volume":"55","author":"Kopylov","year":"2019","journal-title":"Appl. Biochem. Microbiol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"336","DOI":"10.1128\/jcm.33.2.336-341.1995","article-title":"Detection of Yersinia pestis fraction 1 antigen with a fiber optic biosensor","volume":"33","author":"Cao","year":"1995","journal-title":"J. Clin. Microbiol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"572","DOI":"10.1016\/S0035-9203(98)90923-3","article-title":"F1 antigenemia in bubonic plague patients, a marker of gravity and efficacy of treatment","volume":"92","author":"Chanteau","year":"1998","journal-title":"Trans. R Soc. Trop. Med. Hyg."},{"key":"ref_32","first-page":"339","article-title":"LcrV capture enzyme-linked immunosorbent assay for detection of Yersinia pestis from human samples","volume":"12","author":"Savitt","year":"2005","journal-title":"Clin. Diagn. Lab. Immunol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2234","DOI":"10.1128\/IAI.71.4.2234-2238.2003","article-title":"Synergistic protection of mice against plague with monoclonal antibodies specific for the F1 and V antigens of Yersinia pestis","volume":"71","author":"Hill","year":"2003","journal-title":"Infect. Immun."},{"key":"ref_34","first-page":"211","article-title":"Development and testing of an enzyme immunoassay-based monoclonal test system for the detection of the Yersinia pestis V antigen","volume":"50","author":"Ivashchenko","year":"2014","journal-title":"Prikl. Biokhimiia Mikrobiol."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Lillo, A.M., Ayriss, J.E., Shou, Y., Graves, S.W., Bradbury, A.R., and Pavlik, P. (2011). Development of phage-based single chain Fv antibody reagents for detection of Yersinia pestis. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0027756"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1870","DOI":"10.1016\/j.vaccine.2009.11.076","article-title":"Amino acid residues 196-225 of LcrV represent a plague protective epitope","volume":"28","author":"Quenee","year":"2010","journal-title":"Vaccine"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/j.bios.2014.09.090","article-title":"Kinetic epitope mapping of monoclonal antibodies raised against the Yersinia pestis virulence factor LcrV","volume":"65","author":"Read","year":"2015","journal-title":"Biosens. Bioelectron."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"3404","DOI":"10.1128\/JCM.00458-07","article-title":"Comparison of hand-held test kits, immunofluorescence microscopy, enzyme-linked immunosorbent assay, and flow cytometric analysis for rapid presumptive identification of Yersinia pestis","volume":"45","author":"Tomaso","year":"2007","journal-title":"J. Clin. Microbiol."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Xiao, X., Zhu, Z., Dankmeyer, J.L., Wormald, M.M., Fast, R.L., Worsham, P.L., Cote, C.K., Amemiya, K., and Dimitrov, D.S. (2010). Human anti-plague monoclonal antibodies protect mice from Yersinia pestis in a bubonic plague model. PLoS ONE, 5.","DOI":"10.1371\/journal.pone.0013047"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1016\/S0140-6736(03)12270-2","article-title":"Development and testing of a rapid diagnostic test for bubonic and pneumonic plague","volume":"361","author":"Chanteau","year":"2003","journal-title":"Lancet"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1534","DOI":"10.1128\/CVI.00510-14","article-title":"Characterization and epitope mapping of the polyclonal antibody repertoire elicited by ricin holotoxin-based vaccination","volume":"21","author":"Cohen","year":"2014","journal-title":"Clin. Vaccine Immunol."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Noy-Porat, T., Rosenfeld, R., Ariel, N., Epstein, E., Alcalay, R., Zvi, A., Kronman, C., Ordentlich, A., and Mazor, O. (2016). Isolation of Anti-Ricin protective antibodies exibiting high affinity from immunized non-human primates. Toxins, 8.","DOI":"10.3390\/toxins8030064"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Mechaly, A., Alcalay, R., Noy-Porat, T., Epstein, E., Gal, Y., and Mazor, O. (2018). Novel Phage Display-Derived Anti-Abrin Antibodies Confer Post-Exposure Protection against Abrin Intoxication. Toxins, 10.","DOI":"10.3390\/toxins10020080"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1016\/j.mimet.2006.08.004","article-title":"Magnetic biosensor for the detection of Yersinia pestis","volume":"68","author":"Meyer","year":"2007","journal-title":"J. Microbiol. Methods"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1016\/j.jmmm.2006.10.1180","article-title":"New type of biosensor based on magnetic nanoparticle detection","volume":"311","author":"Nikitin","year":"2007","journal-title":"J. Magn. Magn. Mater."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"6283","DOI":"10.1021\/acs.analchem.6b00362","article-title":"Highly Stable Lyophilized Homogeneous Bead-Based Immunoassays for On-Site Detection of Bio Warfare Agents from Complex Matrices","volume":"88","author":"Mechaly","year":"2016","journal-title":"Anal. Chem."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"6866","DOI":"10.1016\/j.vaccine.2011.07.059","article-title":"T cells play an essential role in anti-F1 mediated rapid protection against bubonic plague","volume":"29","author":"Levy","year":"2011","journal-title":"Vaccine"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"P98","DOI":"10.1186\/1475-2859-5-S1-P98","article-title":"Production and purification of high molecular weight oligomers of Yersinia pestis F1 capsular antigen released by high cell density culture of recombinant Escherichia coli cells carrying the caf1 operon","volume":"5","author":"Holtzman","year":"2006","journal-title":"Microb. Cell Factories"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"2854","DOI":"10.1128\/iai.63.8.2854-2858.1995","article-title":"Active immunization with recombinant V antigen from Yersinia pestis protects mice against plague","volume":"63","author":"Leary","year":"1995","journal-title":"Infect. Immun."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/S0168-1656(99)00185-6","article-title":"Endotoxin removal from protein solutions","volume":"76","author":"Petsch","year":"2000","journal-title":"J. Biotechnol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1099\/00221287-21-1-135","article-title":"A study on pesticin biosynthesis","volume":"21","author":"Hertman","year":"1959","journal-title":"J. Gen. Microbiol."},{"key":"ref_52","doi-asserted-by":"crossref","unstructured":"Vagima, Y., Zauberman, A., Levy, Y., Gur, D., Tidhar, A., Aftalion, M., Shafferman, A., and Mamroud, E. (2015). Circumventing Y. pestis Virulence by Early Recruitment of Neutrophils to the Lungs during Pneumonic Plague. PLoS Pathog., 11.","DOI":"10.1371\/journal.ppat.1004893"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1099\/00221287-19-2-289","article-title":"Bacteriocin-like material produced by Pasteurella pestis","volume":"19","author":"Hertman","year":"1958","journal-title":"J. Gen. Microbiol."},{"key":"ref_54","first-page":"196","article-title":"Behavior in vivo of strains of Pasteurella pestis in relation to their efficacy as live vaccines","volume":"117","author":"Markenson","year":"1969","journal-title":"Annales de L\u2019institut Pasteur"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"1147","DOI":"10.1093\/infdis\/jiz260","article-title":"Postexposure Administration of a Yersinia pestis Live Vaccine for Potentiation of Second-Line Antibiotic Treatment Against Pneumonic Plague","volume":"220","author":"Zauberman","year":"2019","journal-title":"J. Infect. Dis."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"3114","DOI":"10.1128\/JB.181.10.3114-3122.1999","article-title":"Molecular characterization of KatY (antigen 5), a thermoregulated chromosomally encoded catalase-peroxidase of Yersinia pestis","volume":"181","author":"Garcia","year":"1999","journal-title":"J. Bacteriol."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"353","DOI":"10.1111\/j.1863-2378.2007.01072.x","article-title":"Mastitis caused by Yersinia pseudotuberculosis in Israeli dairy cattle and public health implications","volume":"54","author":"Shwimmer","year":"2007","journal-title":"Zoonoses Public Health"},{"key":"ref_58","doi-asserted-by":"crossref","unstructured":"Saraka, D., Savin, C., Kouassi, S., Ciss\u00e9, B., Koffi, E., Cabanel, N., Br\u00e9mont, S., Faye-Kette, H., Dosso, M., and Carniel, E. (2017). Yersinia enterocolitica, a Neglected Cause of Human Enteric Infections in C\u00f4te d\u2019Ivoire. PLoS Negl. Trop. Dis., 11.","DOI":"10.1371\/journal.pntd.0005216"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"312","DOI":"10.1007\/978-0-387-72124-8_28","article-title":"Disparity between Yersinia pestis and Yersinia enterocolitica O:8 in YopJ\/YopP-dependent functions","volume":"603","author":"Zauberman","year":"2007","journal-title":"Adv. Exp. Med. Biol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"495","DOI":"10.1038\/256495a0","article-title":"Continuous cultures of fused cells secreting antibody of predefined specificity","volume":"256","author":"Kohler","year":"1975","journal-title":"Nature"},{"key":"ref_61","doi-asserted-by":"crossref","unstructured":"Shafferman, A., Ordentlich, A., and Velan, B. (2010). Protection Against Plague Afforded by Treatment with Polyclonal \u03b1LcrV and \u03b1F1 Antibodies. The Challenge of Highly Pathogenic Microorganism, Springer.","DOI":"10.1007\/978-90-481-9054-6"},{"key":"ref_62","doi-asserted-by":"crossref","unstructured":"Mechaly, A., Vitner, E., Levy, H., Weiss, S., Bar-David, E., Gur, D., Koren, M., Cohen, H., Cohen, O., and Mamroud, E. (2018). Simultaneous immunodetection of anthrax, plague and tularemia from blood cultures utilizing multiplexed suspension arrays. J. Clin. Microbiol.","DOI":"10.1128\/JCM.01479-17"},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"502","DOI":"10.1021\/bi101059w","article-title":"Direct measurement of thermal stability of expressed CCR5 and stabilization by small molecule ligands","volume":"50","author":"Knepp","year":"2011","journal-title":"Biochemistry"},{"key":"ref_64","unstructured":"(1996). ICH Guideline ICHHT Validation of Analytical Procedures: Methodology, EMEA."}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/10\/3\/285\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:31:35Z","timestamp":1760160695000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/10\/3\/285"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,3,2]]},"references-count":64,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2021,3]]}},"alternative-id":["pathogens10030285"],"URL":"https:\/\/doi.org\/10.3390\/pathogens10030285","relation":{},"ISSN":["2076-0817"],"issn-type":[{"value":"2076-0817","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,3,2]]}}}