Physical and chemical characterization of a Giardia lamblia-specific antigen useful in the coprodiagnosis of giardiasis

Abstract

We recently reported the isolation and identification of a Giardia lamblia-specific antigen (GSA 65) that is shed in the stool of giardiasis patients. In the present study, this antigen was affinity purified from sonic extracts of axenically cultured G. lamblia trophozoites and characterized to better understand its biological function and its potential usefulness in the design of coprodiagnostic assays for giardiasis. GSA 65 was resistant to proteolytic digestion with trypsin, chymotrypsin, and protease but was sensitive to treatment with NaIO4 as assessed by Western blotting. This antigen was also stable during prolonged storage at 4 and -20 degrees C in 10% Formalin or distilled H2O as assessed by counterimmunoelectrophoresis. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and isoelectric focusing gel banding patterns, in conjunction with protein and carbohydrate assays and lectin binding studies, confirmed that this antigen is a highly glycosylated glycoprotein. The resistance of GSA 65 to proteolytic degradation, together with previous immunofluorescence data that indicate the antigen is an integral part of the G. lamblia cyst wall, suggests that this molecule may play a role in maintaining the integrity of the cyst in vivo. The ability of GSA 65 to maintain its antigenic structure under a wide variety of conditions makes it an ideal antigen around which to design sensitive immunodiagnostic assays for giardiasis.