Recombinant plasmodium falciparum merozoite surface proteins 4 and 5 and their use
AssigneesInstitut Pasteur, Centre Nationale de la Recherche Scientifique
InventorsShirley Longacre, Hannah Polson, Ronald Perraut, Faridabano Nato
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Abstract
Accordingly, the invention provides constructs in which the nucleic acids encoding Plasmodium falciparum MSP4 and MSP5, and the resulting polypeptides, have been modified. More particularly, this invention provides constructs encoding recombinant MSP4 and MSP5 polypeptides, which are expressed as soluble, secreted polypeptides in a baculovirus-insect cell expression system. It was surprisingly found that the recombinant polypeptides contain an EGF-like domain at the C-terminus that is properly folded in the polypeptide.
Claims63
A purified nucleic acid molecule comprising the DNA sequence of SEQ ID NOS:
- 1, 2, 3, 4, 5, 6, 7, 28, 29, or 8.
A purified nucleic acid molecule encoding an amino acid sequence comprising the sequence of SEQ ID NOS:
- 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 30, or 22.
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A purified nucleic acid molecule, which encodes recombinant MSP4 or MSP5 polypeptide, an allelic variant of recombinant MSP4 or MSP5 polypeptide DNA, or a homolog of recombinant MSP4 or MSP5 polypeptide DNA.
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A recombinant vector selected from the group consisting of I-3512, I-3513, I-3514, I-3694, I-3695 and I-3696.
An immunological complex comprising a recombinant MSP4 and MSP5 polypeptide and an antibody that specifically recognizes said polypeptide.
A method for detecting infection by malaria parasite, wherein the method comprises providing a composition comprising a biological material suspected of being infected with malaria parasite, and assaying for the presence of native MSP4 and/or MSP5 polypeptide of malaria parasite that binds to an antibody specific for recombinant MSP4 or MSP5 polypeptide.
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An in vitro diagnostic method for the detection of the presence or absence of antibodies, which bind to an antigen comprising recombinant MSP4 or MSP5 polypeptide, wherein the method comprises contacting the antigen with a biological fluid for a time and under conditions sufficient for the antigen and antibodies in the biological fluid to form an antigen-antibody complex, and detecting the formation of the complex.
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A diagnostic kit for the detection of the presence or absence of antibodies, which bind to recombinant MSP4 or MSP5 polypeptide or mixtures thereof, wherein the kit comprises an antigen comprising recombinant MSP4 or MSP5 polypeptide or mixtures of recombinant MSP4 and MSP5 polypeptides, and means for detecting the formation of immune complex between the antigen and antibodies, wherein the means are present in an amount sufficient to perform said detection.
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A polynucleotide selected from SEQ ID NO:
- 1, 2, 3, 4, 5, 6, 7, or 8.
A polynucleotide selected from SEQ ID NO:
- 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, or 30.
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A method for in vitro diagnosing a Plasmodium infection in a sample of biological material from a human suspected to be infected by the parasite, wherein the method comprises the following steps:
- contacting an antigen comprising recombinant MSP4 or MSP5 polypeptide with the sample for a time and under conditions sufficient for the antigen and antibodies in the sample to form an antigen-antibody complex, and
- detecting the formation of the complex.
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Hybridoma deposited at C.N.C.M. on Nov. 16, 2005 under Accession number I-3517, I-3518, I-3519 or I-3520.
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Dependencies are inferred from the claim text.