PMID- 31997109 OWN - NLM STAT- MEDLINE DCOM- 20201125 LR - 20231103 IS - 1432-0614 (Electronic) IS - 0175-7598 (Print) IS - 0175-7598 (Linking) VI - 104 IP - 6 DP - 2020 Mar TI - Functional characterization of a broad and potent neutralizing monoclonal antibody directed against outer membrane protein (OMP) of Salmonella typhimurium. PG - 2651-2661 LID - 10.1007/s00253-020-10394-5 [doi] AB - In the present study, we have generated a murine monoclonal antibody (mAb) named Sal-06 by using the crude outer membrane protein preparation of Salmonella enteric subsp. enterica serovar Typhimurium ATCC 14028 strain as antigen. Sal-06mAb belonging to IgG1 isotype demonstrated broad cross-reactivity to standard and isolated strains of genus Salmonella and others such as Escherichia coli, Klebsiella pneumonia, and Proteus mirabilis. Cross-reactivity across several bacterial genera indicated that the epitopes reactive to Sal-06mAb are conserved among these members. Neutralizing effects of Sal-06mAb on Salmonella growth and survival was evaluated in vitro using bacteriostatic and bactericidal activity with and without complement and bacterial invasion inhibition assay. Sal-06mAb demonstrated a bacteriostatic effect on the growth of S. typhimurium ATCC 14028 strain which is both time and concentration (of mAb) dependent. It was also found that the bacterial growth inhibition was complement independent. When the bacterial cells were preincubated with Sal-06mAb, it reduced the adherence and invasion of bacterial cells into A549 epithelial cell line. This was confirmed by CFU count analysis, phase contrast, and fluorescence microscopy. Scanning electron microscope (SEM) imaging confirmed the antimicrobial effects of Sal-06mAb on S. typhimurium ATCC 14028. The development of broadly reactive and cross protective Sal-06mAb opens new possibilities for immunotherapy of sepsis caused by Gram-negative Enterobacteriaceae members. FAU - Reddy, Prakash Narayana AU - Reddy PN AD - Department of Biotechnology, Vignan's Foundation for Science, Technology and Research (Deemed to be University), Vadlamudi, Guntur district, Andhra Pradesh, 522 213, India. FAU - Makam, Shivakiran Sathyanarayan AU - Makam SS AUID- ORCID: 0000-0003-3844-9770 AD - Department of Biotechnology, Vignan's Foundation for Science, Technology and Research (Deemed to be University), Vadlamudi, Guntur district, Andhra Pradesh, 522 213, India. shivasaimsk@gmail.com. FAU - Kota, Rohini Krishna AU - Kota RK AD - Department of Biotechnology, Vignan's Foundation for Science, Technology and Research (Deemed to be University), Vadlamudi, Guntur district, Andhra Pradesh, 522 213, India. FAU - Yatung, Gyati AU - Yatung G AD - Department of Microbiology, Defence Food Research Laboratory (DFRL - DRDO), Siddartha Nagar, Mysore, Karnataka, 570 011, India. FAU - Urs, Radhika Madam AU - Urs RM AD - Department of Microbiology, Defence Food Research Laboratory (DFRL - DRDO), Siddartha Nagar, Mysore, Karnataka, 570 011, India. FAU - Batra, Harshvardhan AU - Batra H AD - Department of Microbiology, Defence Food Research Laboratory (DFRL - DRDO), Siddartha Nagar, Mysore, Karnataka, 570 011, India. FAU - Tuteja, Urmil AU - Tuteja U AD - Department of Microbiology, Defence Research & Development Establishment (DRDE - DRDO), Gwalior, Madhya Pradesh, 474 002, India. LA - eng GR - ECR/2016/000685/DST-SERB, Department of Science and Technology, Government of INDIA/ GR - DST/INSPIRE/04/2017/000565/INSPIRE, DEPARTMENT OF SCIENCE AND TECHNOLOGY, GOVERNMENT OF INDIA/ GR - 09/1253(0002)/2019-EMR-I/Council of Scientific and Industrial Research (CSIR), Government of India/ PT - Journal Article DEP - 20200129 PL - Germany TA - Appl Microbiol Biotechnol JT - Applied microbiology and biotechnology JID - 8406612 RN - 0 (Antibodies, Monoclonal) RN - 0 (Antibodies, Neutralizing) RN - 0 (Antigens, Bacterial) RN - 0 (Bacterial Outer Membrane Proteins) RN - 9007-36-7 (Complement System Proteins) SB - IM MH - A549 Cells MH - Animals MH - Antibodies, Monoclonal/*immunology MH - Antibodies, Neutralizing/*immunology MH - Antigens, Bacterial/immunology MH - Bacterial Adhesion MH - Bacterial Outer Membrane Proteins/*immunology MH - Complement System Proteins MH - Cross Reactions MH - Enterobacteriaceae/immunology MH - Escherichia coli/immunology MH - Female MH - Humans MH - Mice MH - Mice, Inbred BALB C MH - Salmonella typhimurium/*immunology PMC - PMC7080182 OTO - NOTNLM OT - Bactericidal assay OT - Bacteriostatic OT - Cross-reactivity OT - Enterobacteriaceae OT - Monoclonal antibody OT - Neutralizing antibody COIS- The authors declare that they have no conflict of interest. EDAT- 2020/01/31 06:00 MHDA- 2020/11/26 06:00 PMCR- 2020/03/18 CRDT- 2020/01/31 06:00 PHST- 2019/10/09 00:00 [received] PHST- 2020/01/19 00:00 [accepted] PHST- 2020/01/10 00:00 [revised] PHST- 2020/01/31 06:00 [pubmed] PHST- 2020/11/26 06:00 [medline] PHST- 2020/01/31 06:00 [entrez] PHST- 2020/03/18 00:00 [pmc-release] AID - 10.1007/s00253-020-10394-5 [pii] AID - 10394 [pii] AID - 10.1007/s00253-020-10394-5 [doi] PST - ppublish SO - Appl Microbiol Biotechnol. 2020 Mar;104(6):2651-2661. doi: 10.1007/s00253-020-10394-5. Epub 2020 Jan 29.