1. Anti-peptide antibodies differentiate between plasmodial lactate dehydrogenases.
- Author
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Hurdayal R, Achilonu I, Choveaux D, Coetzer TH, and Dean Goldring JP
- Subjects
- Amino Acid Sequence, Animals, Antibodies, Protozoan isolation & purification, Antigens, Protozoan genetics, Chickens, Enzyme-Linked Immunosorbent Assay methods, Humans, Lactate Dehydrogenases genetics, Malaria diagnosis, Malaria enzymology, Malaria immunology, Mice, Models, Molecular, Molecular Sequence Data, Peptides genetics, Protein Conformation, Recombinant Proteins genetics, Recombinant Proteins immunology, Sequence Alignment, Sequence Homology, Amino Acid, Antibodies, Protozoan immunology, Antigens, Protozoan metabolism, Lactate Dehydrogenases immunology, Peptides immunology, Plasmodium enzymology, Plasmodium immunology
- Abstract
Malaria lactate dehydrogenase, a glycolytic enzyme, is a malaria diagnostic target in lateral flow immunochromatographic rapid diagnostic tests. Recombinant Plasmodium yoelii LDH was cloned into the pET-28a vector, expressed and the expressed protein purified from a Ni-NTA affinity matrix. A pan-malarial LDH antibody directed against a common malaria LDH peptide (APGKSDKEWNRDDLL) and two anti-peptide antibodies, each targeting a unique Plasmodium falciparum (LISDAELEAIFDC) and Plasmodium vivax (KITDEEVEGIFDC) LDH peptide were raised in chickens. The antibodies were affinity purified with the appropriate peptide affinity matrix. The affinity purified anti-peptide antibodies detected recombinant P. falciparum, P. vivax and P. yoelii LDH and native P. falciparum and P. yoelii LDH in western blots and immunofluorescence studies. The pan-malarial antibody detected LDH from the three malaria species in western blots. The species-specific anti-peptide antibodies differentiated between P. falciparum and P. vivax LDH. Affinity purified chicken antibodies against recombinant PfLDH, PvLDH and PyLDH proteins each detected the parent and orthologous proteins with similar titers in an ELISA. The study supports an anti-peptide antibody approach to the development of diagnostic reagents., (Copyright (c) 2010 Elsevier Inc. All rights reserved.)
- Published
- 2010
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