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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Main Authors: Seenichamy, Arivudainambi, Bahaman, Abdul Rani, Abdul Mutalib, Abdul Rahim, Bejo, Siti Khairani
Format: Article
Language:English
Published: Hindawi Publishing Corporation 2014
Online Access:http://psasir.upm.edu.my/id/eprint/35072/
http://psasir.upm.edu.my/id/eprint/35072/1/Production%20and%20Characterization%20of%20a%20Polyclonal.pdf
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author Seenichamy, Arivudainambi
Bahaman, Abdul Rani
Abdul Mutalib, Abdul Rahim
Bejo, Siti Khairani
author_facet Seenichamy, Arivudainambi
Bahaman, Abdul Rani
Abdul Mutalib, Abdul Rahim
Bejo, Siti Khairani
author_sort Seenichamy, Arivudainambi
building UPM Institutional Repository
collection Online Access
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.
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spelling upm-350722015-12-29T08:01:20Z http://psasir.upm.edu.my/id/eprint/35072/ Production and characterization of a polyclonal antibody of anti-rLipL21-IgG against Leptospira for early detection of acute leptospirosis Seenichamy, Arivudainambi Bahaman, Abdul Rani Abdul Mutalib, Abdul Rahim Bejo, Siti Khairani 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. Hindawi Publishing Corporation 2014 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/35072/1/Production%20and%20Characterization%20of%20a%20Polyclonal.pdf Seenichamy, Arivudainambi and Bahaman, Abdul Rani and Abdul Mutalib, Abdul Rahim and Bejo, Siti Khairani (2014) Production and characterization of a polyclonal antibody of anti-rLipL21-IgG against Leptospira for early detection of acute leptospirosis. BioMed Research International, 2014. art. no. 592858. pp. 1-8. ISSN 2314-6133; ESSN: 2314-6141 http://www.hindawi.com/journals/bmri/2014/592858/abs/ 10.1155/2014/592858
spellingShingle Seenichamy, Arivudainambi
Bahaman, Abdul Rani
Abdul Mutalib, Abdul Rahim
Bejo, Siti Khairani
Production and characterization of a polyclonal antibody of anti-rLipL21-IgG against Leptospira for early detection of acute leptospirosis
title Production and characterization of a polyclonal antibody of anti-rLipL21-IgG against Leptospira for early detection of acute leptospirosis
title_full Production and characterization of a polyclonal antibody of anti-rLipL21-IgG against Leptospira for early detection of acute leptospirosis
title_fullStr Production and characterization of a polyclonal antibody of anti-rLipL21-IgG against Leptospira for early detection of acute leptospirosis
title_full_unstemmed Production and characterization of a polyclonal antibody of anti-rLipL21-IgG against Leptospira for early detection of acute leptospirosis
title_short Production and characterization of a polyclonal antibody of anti-rLipL21-IgG against Leptospira for early detection of acute leptospirosis
title_sort production and characterization of a polyclonal antibody of anti-rlipl21-igg against leptospira for early detection of acute leptospirosis
url http://psasir.upm.edu.my/id/eprint/35072/
http://psasir.upm.edu.my/id/eprint/35072/
http://psasir.upm.edu.my/id/eprint/35072/
http://psasir.upm.edu.my/id/eprint/35072/1/Production%20and%20Characterization%20of%20a%20Polyclonal.pdf