Cuccurullo Emilia C, Dong Yuemei, Simões Maria L, Dimopoulos George, Bier Ethan
Johns Hopkins University.
University of California, San Diego.
Res Sq. 2023 Dec 19:rs.3.rs-3757253. doi: 10.21203/rs.3.rs-3757253/v1.
Vector control is a crucial strategy for malaria elimination by preventing infection and reducing disease transmission. Most gains have been achieved through insecticide-treated nets (ITNs) and indoor residual spraying (IRS), but the emergence of insecticide resistance among mosquitoes calls for new tools to be applied. Here, we present the development of a highly effective murine monoclonal antibody, targeting the N-terminal region of the gametocyte antigen Pfs230, that can decrease the infection prevalence by > 50% when fed to mosquitoes with gametocytes in an artificial membrane feeding system. We used a standard mouse immunization protocol followed by protein interaction and parasite-blocking validation at three distinct stages of the monoclonal antibody development pipeline: post-immunization, post-hybridoma generation, and final validation of the monoclonal antibody. We evaluated twenty antibodies identifying one (mAb 13G9) with high Pfs230-affinity and parasite-blocking activity. This 13G9 monoclonal antibody could potentially be developed into a transmission-blocking single-chain antibody for expression in transgenic mosquitoes.
病媒控制是通过预防感染和减少疾病传播来消除疟疾的关键策略。大部分成果是通过使用经杀虫剂处理的蚊帐（ITN）和室内滞留喷洒（IRS）取得的，但蚊子中杀虫剂抗性的出现需要应用新工具。在此，我们展示了一种高效鼠单克隆抗体的研发，该抗体靶向配子体抗原Pfs230的N端区域，在人工膜饲血系统中与带有配子体的蚊子一起饲血时，可使感染率降低50%以上。我们采用标准的小鼠免疫方案，随后在单克隆抗体制备流程的三个不同阶段进行蛋白质相互作用和寄生虫阻断验证：免疫后、杂交瘤生成后以及单克隆抗体的最终验证。我们评估了20种抗体，鉴定出一种（单克隆抗体13G9）具有高Pfs230亲和力和寄生虫阻断活性。这种13G9单克隆抗体有可能被开发成一种用于在转基因蚊子中表达的传播阻断单链抗体。
