Verma Reena, Ravichandran Ramakrishnan, Jayaprakash Naatamai S, Kumar Ashok, Vijayalakshmi Mookambeswaran A, Venkataraman Krishnan
Centre for Bio Separation Technology (CBST), VIT University, Vellore, India.
Biotechnol J. 2015 May;10(5):772-82. doi: 10.1002/biot.201400386. Epub 2015 Feb 11.
Adjuvants play an important role in eliciting immune responses and subsequent generation of antibodies with high specificity. Recently, poly(N-isopropylacrylamide) (PNiPAAm), also known as a "smart" polymer, has been proposed as a potential adjuvant for making antibodies and vaccines. This material exhibits efficient delivery, protection against degradation, and preservation of antigen epitopes. In this work, we used both CFA and smart polymer to develop a highly specific murine monoclonal antibody (mAb) against recombinant truncated histidine rich protein2 (HRP2) of Plasmodium falciparum. Our results indicate that the mAbs developed using these adjuvants were able to recognize recombinant HRP2 and native PfHRP2 protein from spent medium. The mAbs generated against recombinant truncated HRP2 showed better sensitivity to the antigen and importantly mAbs generated using PNiPAAm adjuvant were in the range of 10(8)-10(9) M(-1). The mAbs generated using PNiPAAm are very efficient and sensitive in detecting HRP2. To the best of our knowledge, this is the first report of such comparison having been made between these two adjuvants and we propose that the smart polymer has huge potential as an alternative to CFA. Additionally, we discuss the utility of the mAbs generated through PNiPAAm for specific diagnosis of malaria caused by P. falciparum.
佐剂在引发免疫反应以及随后产生高特异性抗体方面发挥着重要作用。最近,聚(N - 异丙基丙烯酰胺)(PNiPAAm),也被称为“智能”聚合物,已被提议作为制备抗体和疫苗的潜在佐剂。这种材料具有高效递送、防止降解以及保留抗原表位的特性。在这项工作中,我们使用弗氏完全佐剂(CFA)和智能聚合物来开发一种针对恶性疟原虫重组截短富含组氨酸蛋白2(HRP2)的高特异性小鼠单克隆抗体(mAb)。我们的结果表明,使用这些佐剂开发的单克隆抗体能够识别来自用过的培养基中的重组HRP2和天然PfHRP2蛋白。针对重组截短HRP2产生的单克隆抗体对抗原表现出更好的敏感性,重要的是,使用PNiPAAm佐剂产生的单克隆抗体的亲和力在10⁸ - 10⁹ M⁻¹范围内。使用PNiPAAm产生的单克隆抗体在检测HRP2方面非常高效且灵敏。据我们所知,这是首次对这两种佐剂进行此类比较的报告,并且我们提出智能聚合物作为CFA的替代品具有巨大潜力。此外,我们还讨论了通过PNiPAAm产生的单克隆抗体在特异性诊断恶性疟原虫引起的疟疾方面的效用。