Production and characterization of a monoclonal antibody against recombinant fatty acid binding protein of Fasciola gigantica.
In Fasciola parasites fatty acid binding proteins (FABPs) are the carrier proteins that help in the uptake of fatty acids from the hosts' fluids. Attempts have been made to utilize both native and recombinant FABP (rFABP) for immunodiagnosis and vaccine development for fasciolosis. In this study, we have produced a number of monoclonal antibodies (MoAbs) against rFABP of Fasciola gigantica. These MoAbs were initially screened against rFABP by ELISA and then tested for their specificities by immunoblotting. Five stable clones were selected and characterized further: four of them were of the isotype IgG(1) while one clone was IgG(2a). All the MoAbs reacted with rFABP which has a molecular weight (MW) of 20 kD and with at least two isoforms of native proteins at MW 14.5 kD that were present in the tegumental antigen (TA) and crude worm extracts, and the excretion-secretion materials. Immunoperoxidase staining of frozen sections of adult parasites by using these MoAbs as primary antibodies indicated that FABP were present in high concentration in the parenchymal cells and reproductive tissues, in low concentration in the tegument and caecal epithelium. All MoAbs cross-reacted with a 14.5 kD antigen present in the whole body (WB) extract of Schistosoma mansoni, while no cross-reactivities were detected with antigens from Eurytrema pancreaticum and Paramphistomum spp.
Sirisriro
A
A
Faculty of Science, Department of Microbiology, Mahidol University, RamaVI Road, Bangkok 10400, Thailand.
Grams
R
R
Vichasri-Grams
S
S
Ardseungneon
P
P
Pankao
V
V
Meepool
A
A
Chaithirayanon
K
K
Viyanant
V
V
Tan-Ariya
P
P
Upatham
E S
ES
Sobhon
P
P
D000818:Animals; D000909:Antibodies, Helminth / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D002352:Carrier Proteins / Q000276:immunology* / Q000302:isolation & purification; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005209:Fasciola / Q000276:immunology*; D005211:Fascioliasis / Q000175:diagnosis / Q000662:veterinary*; D050556:Fatty Acid-Binding Proteins; D015801:Helminth Proteins / Q000276:immunology / Q000302:isolation & purification; D015151:Immunoblotting / Q000662:veterinary; D007124:Immunoenzyme Techniques / Q000662:veterinary; D007074:Immunoglobulin G / Q000276:immunology; D008970:Molecular Weight; D009363:Neoplasm Proteins; D020033:Protein Isoforms; D011994:Recombinant Proteins / Q000276:immunology
D000909:Antibodies, Helminth; D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; D050556:Fatty Acid-Binding Proteins; D015801:Helminth Proteins; D007074:Immunoglobulin G; D009363:Neoplasm Proteins; D020033:Protein Isoforms; D011994:Recombinant Proteins