Kaur, C and Asrith, KP and Ramachandra, SG and Hegde, NR (2024) Immunoinformatics-guided recombinant polypeptide-based enzyme-linked immunosorbent assay for seromonitoring of laboratory animals for minute virus of mice and Kilham rat virus. In: PLoS ONE, 19 (2 Febr).
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Abstract
Subclinical infection of laboratory animals with one or more of several pathogens affects the results of experiments on animals. Monitoring the health of laboratory animals encompasses routine surveillance for pathogens, including several viruses. This study aimed to explore the development of an alternative assay to the existing ones for detecting infection of mice and rats with the parvoviruses minute virus of mice (MVM) and Kilham rat virus (KRV), respectively. Full-length VP2 and NS1 proteins of these parvoviruses, besides fragments containing multiple predicted epitopes stitched together, were studied for serological detection. The optimal dilution of full-length proteins and antigenic regions containing predicted epitopes for coating, test sera, and conjugate was determined using a checkerboard titration at each step. The assays were evaluated vis-à -vis commercially available ELISA kits. The results showed that an engineered fusion of fragments containing multiple predicted MVM VP2 and NS1 epitopes was better than either of the full-length proteins for detecting antibodies in 90 of the tested sera samples. For KRV ELISA, full-length VP2 was better compared to other individual recombinant protein fragments or combinations thereof for the detection of antibodies in sera. This report is the first description of an ELISA for KRV and an improved assay for MVM. Importantly, our assays could be exploited with small volumes of sera. The results also demonstrate the utility of immunoinformatics-driven polypeptide engineering in the development of diagnostic assays and the potential to develop better tests for monitoring the health status of laboratory animals. © 2024 Kaur et al.
| Item Type: | Journal Article |
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| Publication: | PLoS ONE |
| Publisher: | Public Library of Science |
| Additional Information: | The copyright for this article belongs to the Author. |
| Keywords: | alkaline phosphatase; nonstructural protein 1; polypeptide; protein VP2; epitope; recombinant protein, affinity chromatography; animal cell; animal experiment; animal model; antibody titer; antigen binding; antigenicity; Article; asymptomatic infection; B lymphocyte; C6 cell line (glioma); calf (bovine); cell culture; circular dichroism; clinical trial; cloning; dilution; enzyme linked immunosorbent assay; epitope mapping; Escherichia coli; experimental animal; fetal bovine serum; genetic recombination; health status; immunization; immunofluorescence; immunogenicity; immunoinformatics; kilham rat virus; Minute virus of mice; mouse; NIH 3T3 cell line; nonhuman; phenotype; plant gene; polyacrylamide gel electrophoresis; polymerase chain reaction; protein expression; protein purification; rat; recombinant plasmid; sensitivity and specificity; serological surveillance; site directed mutagenesis; skim milk; statistical analysis; virus; virus isolation; virus purification; Western blotting; animal tissue; article; controlled study; diagnosis; ELISA kit; genetic recombination; serological surveillance, Animals; Animals, Laboratory; Antibodies, Viral; Enzyme-Linked Immunosorbent Assay; Epitopes; Immunoinformatics; Mice; Minute Virus of Mice; Parvovirus; Peptides; Rats |
| Department/Centre: | Division of Biological Sciences > Central Animal Facility (Formerly Primate Research Laboratory) |
| Date Deposited: | 05 Apr 2024 11:06 |
| Last Modified: | 05 Apr 2024 11:06 |
| URI: | https://eprints.iisc.ac.in/id/eprint/84704 |
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