Munasinghe Erandi, Athapaththu Maheshi, Abeyewickreme Wimaladharma
Faculty of Medicine, University of Kelaniya, Ragama, Sri Lanka.
Industrial Technology Institute, Colombo, Sri Lanka.
Front Mol Biosci. 2022 Oct 21;9:935456. doi: 10.3389/fmolb.2022.935456. eCollection 2022.
Synthetic peptides have recently become common as antigens for antibody production. Peptides are short chains of amino acids that can be used to elicit an immune response. The immunogenicity of the peptide antigens varies depending on the length, charge, solubility, and amino acids contained in the peptide sequence. Dengue NS1 protein is an important target antigen in the early detection of dengue infection. In this study, peptides corresponding to a highly conserved region from the dengue NS1 region were designed and synthesized. Balb/C mice were immunized against each peptide and spleen cells extracted from the immunized mice were fused with NS0 murine myeloma cells. Hybridoma clones obtained from the fusions were tested against peptides using ELISA. Out of 1,830 growing clones, 28 clones produced antibodies reacting with dengue NS1 peptides. A purified monoclonal antibody reacting with all four peptides was tested for reactivity with dengue NS1 native protein using dengue-confirmed serum and urine samples. The monoclonal antibody shows significant reactivity with both serum and urine. The findings of the current research can be used to detect dengue infection using urine, which ultimately results in the prevention of dengue epidemics through painless diagnosis, following treatment, and patient management to safeguard human and economic wellness.
合成肽最近已成为用于抗体生产的常见抗原。肽是氨基酸的短链，可用于引发免疫反应。肽抗原的免疫原性因肽序列中的长度、电荷、溶解度和所含氨基酸而异。登革热NS1蛋白是登革热感染早期检测中的重要靶抗原。在本研究中，设计并合成了与登革热NS1区域高度保守区域相对应的肽。用每种肽对Balb/C小鼠进行免疫，从免疫小鼠中提取脾细胞并与NS0鼠骨髓瘤细胞融合。使用ELISA对融合得到的杂交瘤克隆进行肽检测。在1830个生长的克隆中，有28个克隆产生了与登革热NS1肽反应的抗体。使用登革热确诊的血清和尿液样本，检测一种与所有四种肽反应的纯化单克隆抗体与登革热NS1天然蛋白的反应性。该单克隆抗体与血清和尿液均表现出显著反应性。当前研究的结果可用于通过尿液检测登革热感染，最终通过无痛诊断、后续治疗和患者管理来预防登革热流行，以保障人类健康和经济福祉。
