Development of Standard and Competitive ELISA for Detection and Quantification of PRP-1 in Biological Fluids by Using anti-PRP-1 Polyclonal Antiserum
| Autor: | N. V. Tumasyan, I. K. Sahakyan, N. V. Kocharyan, A. A. Khachatryan, T. K. Davtyan, K. A. Galoyan, S. S. Abrahamyan |
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| Jazyk: | angličtina |
| Rok vydání: | 2024 |
| Předmět: | |
| Zdroj: | International Journal of Biomedicine, Vol 14, Iss 3, Pp 516-519 (2024) |
| Druh dokumentu: | article |
| ISSN: | 2158-0510 2158-0529 |
| DOI: | 10.21103/Article14(3)_OA21 |
| Popis: | This study aimed to establish a sensitive method for the detection of proline-rich polypeptide-1 (PRP-1) in biological fluids. PRP-1, also known as galarmin, is a fragment of neurophysin-vasopressin-associated glycoprotein synthesized by brain neurosecretory cells and consisting of 15 amino acid residues. An enzyme-linked immunosorbent assay (ELISA) was used for PRP-1 quantification. An ELISA system has been developed using polyclonal antibodies we raised against the synthetic PRP-1. According to the analysis, the concentration PRP-1 of 25 ng/mL was accepted as the main appropriate coating concentration for further experiments in 1:100 and 1:500 antibody dilutions. Then, a competitive ELISA was developed to quantify PRP-1 in the fluids. Based on the results, an appropriate condition was chosen to be the best condition for PRP-1 detection: the appropriate quantity of the immobilized PRP-1 (25 ng/mL); anti-rabbit primary antibodies against PRP-1 (1:500); anti-rabbit secondary antibodies conjugated to peroxidase (1:1000), and extravidin (1:1000); as a result, the minimum detectable amount of PRP-1 in the fluid was 1.5 ng/mL. Thus, this method provides a good detection limit and sensitivity and is easy to use. In addition, a large number of rats and human serum and plasma samples can be analyzed rapidly and simultaneously, which is what we intend to realize in the future. |
| Databáze: | Directory of Open Access Journals |
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