Epitope-based in silico peptide design yields peptide-directed antibodies that recognize the buffalo luteinizing hormone

• Published in: International journal of biological macromolecules Vol. 176; pp. 260 - 271
• Main Authors: Nayan, Varij, Onteru, Suneel Kumar, Singh, Dheer
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 15.04.2021
• Subjects: Anti-peptide antibodies; Luteinizing hormone; Water buffalo; Luteinizing hormone; Anti-peptide antibodies; Water buffalo
• ISSN: 0141-8130; 1879-0003
• DOI: 10.1016/j.ijbiomac.2021.02.083

We present a novel peptide sequence identified through in silico epitope design and the later generation of peptide-directed antibodies recognizing the buffalo luteinizing hormone. Peptides and antibodies, specific to reproductive hormones, are valuable tools for developing point-of-care immunodiagnostic tools. The study predicted an epitope peptide in silico from buffalo luteinizing hormone and the generation of polyclonal antibodies against this peptide sequence. In this quest, we identified a novel epitope peptide sequence (luteinizing hormone peptide, LHP) through bioinformatics tools. The peptide was further synthesized and characterized. The polyclonal antibodies (anti-LHP) were raised against the peptide in the rabbit. Thereafter, we explored a strategy for detecting buffalo luteinizing hormone (LH) using the anti-peptide antibodies developed. The affinity of the peptide, bovine lutropin beta, and crude LH (prepared from buffalo pituitary) towards the raised antibodies was established by dot blot and ELISA. Specific recognition of the luteinizing hormone by the raised polyclonal antibodies highlights the ability of the identified peptide (LHP) and developed polyclonal antibodies (anti-LHP) as suitable diagnostic reagents for sensing the buffalo luteinizing hormone. Through this work, we analyzed and translated the “-omics” information in the LH gene sequence for the development of a novel peptide and antibodies as valuable immuno-reagents. [Display omitted] •A novel epitope was identified in silico from the buffalo LH beta subunit.•The functionality of the polyclonal antibodies was verified against the epitope peptide and the buffalo LH.•The affinity of LHP, lutropin beta, and crude LH was established towards raised anti-LHP.•The peptide and developed polyclonal antibodies can serve as diagnostic reagents.