Production and characterization of immunoglobulin G anti-rLipL32 antibody as a biomarker for the diagnosis of leptospirosis

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

Susanti Susanti¹, Pratiwi Pudjilestari Sudarmono², N. L. P. Indi Dharmayanti³, and Prasandhya Astagiri Yusuf⁴

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: Accepted: Published online:

Corresponding author: Susanti Susanti E-mail: santibadgi@gmail.com

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.