Adjuvant poly(N‐isopropylacrylamide) generates more efficient monoclonal antibodies against truncated recombinant histidine‐rich protein2 of Plasmodium falciparum for malaria diagnosis

Abstract

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.

Diagnostic potential of monoclonal antibodies developed against C-terminal polypeptide of P. falciparum Histidine Rich Protein2 (PfHRP2) in malaria infected patients from India

“…Importantly, this uniqueness is believed to be conserved in different P. falciparum isolates originating from various parts of the world. So far no attempts have been made to develop mAbs against this unique region comprising of various peptide repeats of PfHRP2 [12,13].…”

“…The recP-fHRP2-T3 protein was used as an antigen for the development of both polyclonal (in laboratory-bred female New Zealand rabbits) and monoclonal antibody (6-12 weeks old female BALB/c mice). Two clones from the mAbs produced (b10c1 and Aa3c10) having affinity constant of 10 9 M −1 (determined by Surface Plasma Resonance using Ni-NTA column) were very promising [12,13]. Purified mAbs and polyclonal antibodies, were evaluated with malaria infected patient samples.…”