Production and Characterization of a Polyclonal Antibody of Anti-rLipL21-IgG against Leptospira for Early Detection of Acute Leptospirosis
Leptospirosis is one of the zoonotic diseases in animals and humans throughout the world. LipL21 is one of the important surface-exposed lipoproteins in leptospires and the most effective cross protective immunogenic antigen. It is widely considered as a diagnostic marker for leptospirosis. In this...
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| creator | Seenichamy, Arivudainambi Bahaman, Abdul Rani Mutalib, Abdul Rahim Khairani-Bejo, Siti |
| description | Leptospirosis is one of the zoonotic diseases in animals and humans throughout the world. LipL21 is one of the important surface-exposed lipoproteins in leptospires and the most effective cross protective immunogenic antigen. It is widely considered as a diagnostic marker for leptospirosis. In this study, we evaluated the serodiagnostic potential of LipL21 protein of Leptospira interrogans serovar Pomona. We have successfully amplified, cloned, and expressed LipL21 in E. coli and evaluated its specificity by immunoblotting. Purified recombinant LipL21 (rLipL21) was inoculated into rabbits for the production of polyclonal antibody. Characterization of the purified IgG antibody against rLipL21 was performed by cross reactivity assay. Only sera from leptospirosis patients and rabbit hyperimmune sera recognized rLipL21 while the nonleptospirosis control sera showed no reaction in immunoblotting. We confirmed that anti-rLipL21-IgG antibody cross reacted with and detected only pathogenic leptospiral species and it did not react with nonpathogenic leptospires and other bacterial species. Results observed showed that anti-rLipL21-IgG antibody has high specificity and sensitivity to leptospires. The findings indicated that the antibody could be used in a diagnostic assay for detection of leptospires or their proteins in the early phase of infection. |
| doi_str_mv | 10.1155/2014/592858 |
| format | Article |
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LipL21 is one of the important surface-exposed lipoproteins in leptospires and the most effective cross protective immunogenic antigen. It is widely considered as a diagnostic marker for leptospirosis. In this study, we evaluated the serodiagnostic potential of LipL21 protein of Leptospira interrogans serovar Pomona. We have successfully amplified, cloned, and expressed LipL21 in E. coli and evaluated its specificity by immunoblotting. Purified recombinant LipL21 (rLipL21) was inoculated into rabbits for the production of polyclonal antibody. Characterization of the purified IgG antibody against rLipL21 was performed by cross reactivity assay. Only sera from leptospirosis patients and rabbit hyperimmune sera recognized rLipL21 while the nonleptospirosis control sera showed no reaction in immunoblotting. We confirmed that anti-rLipL21-IgG antibody cross reacted with and detected only pathogenic leptospiral species and it did not react with nonpathogenic leptospires and other bacterial species. Results observed showed that anti-rLipL21-IgG antibody has high specificity and sensitivity to leptospires. The findings indicated that the antibody could be used in a diagnostic assay for detection of leptospires or their proteins in the early phase of infection.</description><identifier>ISSN: 2314-6133</identifier><identifier>EISSN: 2314-6141</identifier><identifier>DOI: 10.1155/2014/592858</identifier><identifier>PMID: 24860824</identifier><language>eng</language><publisher>Cairo, Egypt: Hindawi Puplishing Corporation</publisher><subject>Antibodies, Monoclonal - biosynthesis ; Antibodies, Monoclonal - immunology ; Antigens, Bacterial - immunology ; Bacteriology ; Care and treatment ; Chemical properties ; Cloning ; Deoxyribonucleic acid ; DNA ; E coli ; Escherichia coli ; Health aspects ; Humans ; Immunoassay - methods ; Immunoglobulin G - immunology ; Infections ; Leptospira - immunology ; Leptospira interrogans ; Leptospirosis ; Leptospirosis - diagnosis ; Leptospirosis - immunology ; Leptospirosis - microbiology ; Lipoproteins ; Proteins ; Reproducibility of Results ; Ribosomal Proteins - immunology ; Sensitivity and Specificity ; Serologic Tests - methods ; Tropical diseases ; Veterinary medicine</subject><ispartof>BioMed research international, 2014-01, Vol.2014 (2014), p.1-8</ispartof><rights>Copyright © 2014 Arivudainambi Seenichamy et al.</rights><rights>COPYRIGHT 2014 John Wiley & Sons, Inc.</rights><rights>Copyright © 2014 Arivudainambi Seenichamy et al. Arivudainambi Seenichamy et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</rights><rights>Copyright © 2014 Arivudainambi Seenichamy et al. 2014</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c490t-ed4cdfaf982c1ac70a74e6d14dbab5d34f7c996fcb74822049603346c7edfd0a3</citedby><cites>FETCH-LOGICAL-c490t-ed4cdfaf982c1ac70a74e6d14dbab5d34f7c996fcb74822049603346c7edfd0a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4016889/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4016889/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,315,728,781,785,886,27929,27930,53796,53798</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24860824$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Soares, Marcelo A.</contributor><creatorcontrib>Seenichamy, Arivudainambi</creatorcontrib><creatorcontrib>Bahaman, Abdul Rani</creatorcontrib><creatorcontrib>Mutalib, Abdul Rahim</creatorcontrib><creatorcontrib>Khairani-Bejo, Siti</creatorcontrib><title>Production and Characterization of a Polyclonal Antibody of Anti-rLipL21-IgG against Leptospira for Early Detection of Acute Leptospirosis</title><title>BioMed research international</title><addtitle>Biomed Res Int</addtitle><description>Leptospirosis is one of the zoonotic diseases in animals and humans throughout the world. LipL21 is one of the important surface-exposed lipoproteins in leptospires and the most effective cross protective immunogenic antigen. It is widely considered as a diagnostic marker for leptospirosis. In this study, we evaluated the serodiagnostic potential of LipL21 protein of Leptospira interrogans serovar Pomona. We have successfully amplified, cloned, and expressed LipL21 in E. coli and evaluated its specificity by immunoblotting. Purified recombinant LipL21 (rLipL21) was inoculated into rabbits for the production of polyclonal antibody. Characterization of the purified IgG antibody against rLipL21 was performed by cross reactivity assay. Only sera from leptospirosis patients and rabbit hyperimmune sera recognized rLipL21 while the nonleptospirosis control sera showed no reaction in immunoblotting. We confirmed that anti-rLipL21-IgG antibody cross reacted with and detected only pathogenic leptospiral species and it did not react with nonpathogenic leptospires and other bacterial species. Results observed showed that anti-rLipL21-IgG antibody has high specificity and sensitivity to leptospires. The findings indicated that the antibody could be used in a diagnostic assay for detection of leptospires or their proteins in the early phase of infection.</description><subject>Antibodies, Monoclonal - biosynthesis</subject><subject>Antibodies, Monoclonal - immunology</subject><subject>Antigens, Bacterial - immunology</subject><subject>Bacteriology</subject><subject>Care and treatment</subject><subject>Chemical properties</subject><subject>Cloning</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>E coli</subject><subject>Escherichia coli</subject><subject>Health aspects</subject><subject>Humans</subject><subject>Immunoassay - methods</subject><subject>Immunoglobulin G - immunology</subject><subject>Infections</subject><subject>Leptospira - immunology</subject><subject>Leptospira interrogans</subject><subject>Leptospirosis</subject><subject>Leptospirosis - diagnosis</subject><subject>Leptospirosis - immunology</subject><subject>Leptospirosis - microbiology</subject><subject>Lipoproteins</subject><subject>Proteins</subject><subject>Reproducibility of Results</subject><subject>Ribosomal Proteins - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>BioMed research international</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Seenichamy, Arivudainambi</au><au>Bahaman, Abdul Rani</au><au>Mutalib, Abdul Rahim</au><au>Khairani-Bejo, Siti</au><au>Soares, Marcelo A.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Production and Characterization of a Polyclonal Antibody of Anti-rLipL21-IgG against Leptospira for Early Detection of Acute Leptospirosis</atitle><jtitle>BioMed research international</jtitle><addtitle>Biomed Res Int</addtitle><date>2014-01-01</date><risdate>2014</risdate><volume>2014</volume><issue>2014</issue><spage>1</spage><epage>8</epage><pages>1-8</pages><issn>2314-6133</issn><eissn>2314-6141</eissn><abstract>Leptospirosis is one of the zoonotic diseases in animals and humans throughout the world. LipL21 is one of the important surface-exposed lipoproteins in leptospires and the most effective cross protective immunogenic antigen. It is widely considered as a diagnostic marker for leptospirosis. In this study, we evaluated the serodiagnostic potential of LipL21 protein of Leptospira interrogans serovar Pomona. We have successfully amplified, cloned, and expressed LipL21 in E. coli and evaluated its specificity by immunoblotting. Purified recombinant LipL21 (rLipL21) was inoculated into rabbits for the production of polyclonal antibody. Characterization of the purified IgG antibody against rLipL21 was performed by cross reactivity assay. Only sera from leptospirosis patients and rabbit hyperimmune sera recognized rLipL21 while the nonleptospirosis control sera showed no reaction in immunoblotting. We confirmed that anti-rLipL21-IgG antibody cross reacted with and detected only pathogenic leptospiral species and it did not react with nonpathogenic leptospires and other bacterial species. Results observed showed that anti-rLipL21-IgG antibody has high specificity and sensitivity to leptospires. The findings indicated that the antibody could be used in a diagnostic assay for detection of leptospires or their proteins in the early phase of infection.</abstract><cop>Cairo, Egypt</cop><pub>Hindawi Puplishing Corporation</pub><pmid>24860824</pmid><doi>10.1155/2014/592858</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Antibodies, Monoclonal - biosynthesis Antibodies, Monoclonal - immunology Antigens, Bacterial - immunology Bacteriology Care and treatment Chemical properties Cloning Deoxyribonucleic acid DNA E coli Escherichia coli Health aspects Humans Immunoassay - methods Immunoglobulin G - immunology Infections Leptospira - immunology Leptospira interrogans Leptospirosis Leptospirosis - diagnosis Leptospirosis - immunology Leptospirosis - microbiology Lipoproteins Proteins Reproducibility of Results Ribosomal Proteins - immunology Sensitivity and Specificity Serologic Tests - methods Tropical diseases Veterinary medicine |
| title | Production and Characterization of a Polyclonal Antibody of Anti-rLipL21-IgG against Leptospira for Early Detection of Acute Leptospirosis |
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