Development of B cell epitopes-based enzyme linked immune sorbent assay for detection of bovine anti-Mullerian hormone

• Published in: 3 Biotech Vol. 13; no. 7; p. 241
• Main Authors: Pal, Prasanna, Aggarwal, Anjali, Rajput, Y. S., Deb, Rajib, Joshi, Vinay G., Verma, Arvind Kumar, Haldar, Avijit, Singh, Indra, Grewal, Sonika, De, Sachinandan
• Format: Journal Article
• Language: English
• Published: Cham Springer International Publishing 01-07-2023; Springer Nature B.V
• Subjects: Agriculture; Antibodies; Assaying; Bioinformatics; Biomaterials; Biotechnology; Cancer Research; Cattle; Chemistry; Chemistry and Materials Science; Enzyme-linked immunosorbent assay; Epitopes; Follicle-stimulating hormone; Luteinizing hormone; Peptides; Progesterone; Protocols and Methods; Recovery; Sorbents; Stem Cells; B cell epitope; Bovine; Antibody; bAMH; Anti-Mullerian hormone; ELISA
• ISSN: 2190-572X; 2190-5738
• DOI: 10.1007/s13205-023-03622-y

The present study aimed to generate antibodies against predicted B cell epitopic peptides encoding bAMH for developing different ELISA models. Sandwich ELISA was determined to be an excellent technique for assessing bAMH in bovine plasma based on sensitivity tests. The assay's specificity, sensitivity, inter- and intra-assay CV, recovery %, Lower limit of quantification (LLOQ), and Upper limit of quantification (ULOQ) were determined. The test was selective since it did not bind to AMH-related growth and differentiation factors (LH and FSH) or non-related components (BSA, progesterone). The intra-assay CV was 5.67%, 3.12%, 4.94%, 3.61% and 4.27% for 72.44, 183.11, 368.24, 522.24 and 732.25 pg/ml AMH levels, respectively. At the same time, the inter-assay CV was 8.77%, 7.87%, 4.53%, 5.76% and 6.70% for 79.30, 161.27, 356.30, 569.33 and 798.19 pg/ml AMH levels, respectively. The average (Mean ± SEM) recovery percentages were 88–100%. LLOQ was 5 pg/ml and ULOQ at 50 µg/ml (CV < 20%). In conclusion, we developed a new highly sensitive ELISA against bAMH using epitope specific antibodies.