Vet World Vol.17 April-2024 Article - 16
Research Article
Veterinary World, 17(4): 871-879
https://doi.org/10.14202/vetworld.2024.871-879
Production and characterization of immunoglobulin G anti-rLipL32 antibody as a biomarker for the diagnosis of leptospirosis
1 Doctoral Program in Biomedical Sciences, Faculty of Medicine, Universitas Indonesia, Jakarta, 10430, Indonesia.
2 Department of Microbiology, Faculty of Medicine, Universitas Indonesia, Jakarta, 10430, Indonesia.
3 Research Center for Veterinary Science, Research Organization for Health, National Research and Innovation Agency, Bogor, 16911, Indonesia.
4 Medical Physiology Biophysics Department and Medical Technology IMERI, Faculty of Medicine, Universitas Indonesia, Jakarta, 10430, Indonesia.
Background and Aim: Microscopic agglutination test (MAT) for the diagnosis of leptospirosis requires live cultures and is serovar-specific, while polymerase chain reaction (PCR) requires expensive equipment and sample preparation. The rLipL32 protein is conserved and can be used for the production of immunoglobulin G (IgG) anti-rLipL32 antibody, which can be used as a biomarker for leptospirosis diagnosis. This study aimed to produce and characterize an IgG anti-rLipL32 antibody as a biomarker for leptospirosis diagnosis.
Materials and Methods: Escherichia coli rLipL32 was cultured and analyzed by PCR and sequencing. Cultures were used for rLipL32 protein expression and purification and the rLipL32 protein was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The rLipL32 protein was used to produce anti-rLipL32 serum and was analyzed by enzyme-linked immunosorbent assay (ELISA). Serum was purified to obtain IgG anti-rLipL32 antibody and characterized by SDS-PAGE and western blotting.
Results: PCR was able to amplify the LipL32 gene from E. coli rLipL32, and sequencing analysis showed 99.19% similarity with pathogenic Leptospira. SDS-PAGE analysis showed a 32-kDa band. ELISA results showed an increase in OD in anti-rLipL32 serum compared to preimmune serum. Western blotting results showed that the IgG anti-rLipL32 antibody was able to bind and cross-reacts with pathogenic Leptospira serovar but not with E. coli or Staphylococcus aureus.
Conclusion: IgG anti-rLipL32 antibody has high specificity and sensitivity against Leptospira pathogens. These findings suggest that IgG anti-rLipL32 antibody is a promising biomarker for the diagnosis of leptospirosis.
Keywords: anti-rLipL32 serum, immunoglobulin G anti-rLipL32 antibody, Leptospira, rLipL32 protein.
How to cite this article: Susanti S, Sudarmono PP, Dharmayanti NLPI, and Yusuf PA (2024) Production and characterization of immunoglobulin G anti-rLipL32 antibody as a biomarker for the diagnosis of leptospirosis, Veterinary World, 17(4): 871-879.
Received: 2023-11-12 Accepted: 2024-03-20 Published online: 2024-04-19
Corresponding author: E-mail:
DOI: 10.14202/vetworld.2024.871-879
Copyright: Susanti, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/ by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/ publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
