Anti-peptide antibodies differentiate between plasmodial lactate dehydrogenases.
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.
Hurdayal
Ramona
R
Biochemistry, University of KwaZulu-Natal, PB X01, Carbis Road, Scottsville 3209, South Africa.
Achilonu
Ikechukwu
I
Choveaux
David
D
Coetzer
Theresa H T
TH
Dean Goldring
J P
JP
D000595:Amino Acid Sequence; D000818:Animals; D000913:Antibodies, Protozoan / Q000276:immunology* / Q000302:isolation & purification; D000953:Antigens, Protozoan / Q000235:genetics / Q000378:metabolism*; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006801:Humans; D045022:Lactate Dehydrogenases / Q000235:genetics / Q000276:immunology*; D008288:Malaria / Q000175:diagnosis / Q000201:enzymology / Q000276:immunology; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010455:Peptides / Q000235:genetics / Q000276:immunology*; D010961:Plasmodium / Q000201:enzymology / Q000276:immunology; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D010455:Peptides; D011994:Recombinant Proteins; D045022:Lactate Dehydrogenases