Immobilization of viral and mycoplasma antigens and of immunoglobulins on polystyrene surface for immunoassays

Abstract

The immobilization of purified influenza virus, rubella virus, crude nuclear cytomegalovirus antigen and of mycoplasma on polystyrene tubes was studied using radio-iodinated preparations. The antigen activities on tube surfaces were determined using sequentially specific human antibodies and alkaline phosphatase-conjugated anti-human IgG in an enzyme-immunoassay (EIA) reaction. In addition, immobilization of radio-iodinated human IgG, IgM, and IgA, serving as model proteins, was studied using the respective anti-immunoglobulin conjugates in EIA directly. Pretreatment of the surface with albumin and glutaraldehyde inhibited the adsorption and antigenicity of IgG. Increase of temperature and thus of speed of adsorption did not affect the fraction of antigen eluted during the test procedure. Only with IgG and IgA was it necessary to saturate the polystyrene surface in order to achieve maximal reactivity in EIA. With other antigens, maximal reactivity in EIA was obtained with amounts of protein much lower than the maximal amount that could be adsorbed per tube. IgM was found to have an exceptionally high affinity to polystyrene.