title: Antigenicity of a Bacterially Expressed Triple Chimeric Antigen of Plasmodium falciparum AARP, MSP-311 and MSP-119: PfAMSP-Fu35.
abstract: Development of fusion chimeras as potential vaccine candidates is considered as an attractive strategy to generate effective immune responses to more than one antigen using a single construct. Here, we described the design, production, purification and antigenicity of a fusion chimera (PfAMSP-Fu35), comprised of immunologically relevant regions of three vaccine target malaria antigens, PfAARP, PfMSP-3 and PfMSP-1. The recombinant PfAMSP-Fu35 is expressed as a soluble protein and purified to homogeneity with ease at a yield of ~ 7 mg L-1. Conformational integrity of the C-terminal fragment of PfMSP-1, PfMSP-119 was retained in the fusion chimera as shown by ELISA with conformation sensitive monoclonal antibodies. High titre antibodies were raised to the fusion protein and to all the three individual components in mice and rabbits upon immunization with fusion chimera in two different adjuvant formulations. The sera against PfAMSP-Fu35 recognized native parasite proteins corresponding to the three components of the fusion chimera. As shown by invasion inhibition assay and antibody mediated cellular inhibition assay, antibodies purified from the PfAMSP-Fu35 immunized serum successfully and efficiently inhibited parasite invasion in P. falciparum 3D7 in vitro both directly and in monocyte dependent manner. However, the invasion inhibitory activity of anti-AMSP-Fu35 antibody is not significantly enhanced as expected as compared to a previously described two component fusion chimera, MSP-Fu24. Therefore, it may not be of much merit to consider AMSP-Fu35 as a vaccine candidate for preclinical development.
authors: Malaria Research Group, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi, India.
authors: Aakanksha
authors: A
authors: Kalra
authors: Malaria Research Group, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi, India.
authors: Jyotheeswara Reddy
authors: JR
authors: Edula
authors: Malaria Research Group, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi, India.
authors: Puneet Kumar
authors: PK
authors: Gupta
authors: Malaria Research Group, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi, India.
authors: Alok Kumar
authors: AK
authors: Pandey
authors: Malaria Research Group, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi, India.
authors: Virander S
authors: VS
authors: Chauhan
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000276:immunology; D000953:Antigens, Protozoan / Q000235:genetics / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D005620:Freund's Adjuvant; D007114:Immunization; D007074:Immunoglobulin G / Q000276:immunology; D017780:Malaria Vaccines / Q000235:genetics / Q000276:immunology; D016778:Malaria, Falciparum / Q000276:immunology / Q000517:prevention & control; D051379:Mice; D009000:Monocytes / Q000276:immunology / Q000378:metabolism; D010963:Plasmodium falciparum / Q000235:genetics / Q000276:immunology*; D011485:Protein Binding; D015800:Protozoan Proteins / Q000235:genetics / Q000276:immunology*; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology*
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D007074:Immunoglobulin G; D017780:Malaria Vaccines; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; D005620:Freund's Adjuvant