Conjugation of coxsackievirus type B1-B6 immunoglobulins with fluorescein isothiocyanate by a "reversed" dialysis method.
Immunoglobulins from antisera to coxsackievirus B types 1--6 were labeled with fluorescein isothiocyanate by a "reverse" dialysis method of conjugation. Conjugates thus obtained were labeled at reproducible ratios of fluorescein to protein with weight-weight ratios ranging from 5.0 to 7.1 and estimated molar ratios ranging from 2.1 to 2.9. Conjugates were tested on cover-slip HeLa cell cultures by the direct method of staining. Two types of specific immunofluorescence were observed: intensely fluorescent perinuclear masses and discrete foci of bright fluorescence scattered throughout the cytoplasm. Titers of homotypic conjugates varied from 1:40 to 1:160. Heterotypic staining, consisting of a diffused hazy fluorescence, was observed only at the highest concentrations of conjugate and not above the 1:10 dilution. This technique proved to be specific and sensitive for the detection of group B coxsackievirus antigens in HeLa cells but failed in specific detection of the same antigens in samples of the myocardium obtained by biopsy of three patients clinically ill with idiopathic myocarditis.
D000914:Antibodies, Viral / Q000032:analysis; D000956:Antigens, Viral / Q000032:analysis*; D004770:Enterovirus / Q000276:immunology*; D005452:Fluoresceins / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D007106:Immune Sera; D013194:Staining and Labeling
D000914:Antibodies, Viral; D000956:Antigens, Viral; D005452:Fluoresceins; D007106:Immune Sera