Biochemical and immunological characterization of soluble antigens of Giardia lamblia.
The crude soluble antigens (CSA) of Giardia lamblia trophozoites and their analytically purified fractions were characterized biochemically and immunologically to determine the most immunogenic fraction and its localization on the parasite. Both Sephacryl S-300 column chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed the highly complex and heterogeneous nature of CSA. Gel filtration of CSA showed four fractions (FI-FIV) with molecular masses of 250, 150, 110, and 10 kDa for fractions I-IV, respectively. Protein profiles of CSA demonstrated 28 Coomassie-blue bands in the range of 200-14 kDa. Similar banding patterns with fewer polypeptides were observed in the FI fraction. However, fractions II and III showed polypeptide bands in the region of 97-14 kDa. The glycoprotein nature of CSA and its-fractions were demonstrated in physicochemical analysis. In antigenic activity analysis the high-molecular-weight FI antigen was found to be 8 times more immunogenic than CSA as well as the other fractions. Major differences in the immunoreactivity of CSA and FI antigens were noted at 220, 30, and 22 kDa for the FI antigen and at 205, 84, 55, 43, and 20 kDa for CSA. Some of these FI polypeptides were found to be surface-associated as revealed by immunofluorescence and immunoblot assay. These results suggest the future use of the FI antigen in the serodiagnosis of and immunoprophylaxis against giardiasis.
Chaudhuri
P P
PP
Department of Parasitology, National Institute of Cholera and Enteric Diseases, Calcutta, India.
Das
D
D
Sarkar
S
S
Munoz
M L
ML
Das
P
P
D000818:Animals; D000953:Antigens, Protozoan / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D015153:Blotting, Western; D003378:Counterimmunoelectrophoresis; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D019084:Fluorescent Antibody Technique, Indirect; D016829:Giardia lamblia / Q000737:chemistry / Q000276:immunology*; D010455:Peptides; D011817:Rabbits; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry
D000953:Antigens, Protozoan; D010455:Peptides; D012967:Sodium Dodecyl Sulfate