PMID: 374631Jan 1, 1979Paper

A new method for fluorescence immunoassay using plane surface solid phases (FIAPS)

Journal of Immunological Methods
H H SedlacekF R Seiler

Abstract

A method distinguished by high sensitivity, low non-specific binding, easy handling, and broad applicability with respect to various antigens is described. Films of polymethyl-methacrylate with plane surfaces were selected as solid phase for adhesive or covalent binding of different antigens (DNA, histone, human, rabbit or goat immunoglobulins). Proteins were covalently bound to the films by the azide method (Orth and Brummer, 1972). Polymethylmethacrylate films thus coated had a negligible autofluorescence and gave minimal non-specific binding of protein. Coated films were used for specificity control of FITC-labeled antibody preparations and in the double antibody and sandwich techniques for detection of antibodies or antigens in sera from man, rabbit and goat. FITC-conjugated hyperimmune antibody, in some cases purified by immunoadsorption was used as second antibody in indirect techniques. The amount of fluorescent-labeled antibody bound per unit of surface area of film was measured by incident light with a Zeiss-Axiomat fluorescence microscope equipped for fluorescence photometry and an uranyl acetate glass plate was used as a standard. The technique appears superior to present methods of quantitative immunofluorescence an...Continue Reading

References

Jun 30, 1975·Annals of the New York Academy of Sciences·W KnappW Hijmans
Aug 12, 1976·Biochimica Et Biophysica Acta·L PecciC Cannella
Nov 1, 1969·Archives of Biochemistry and Biophysics·M Steinbuch, R Audran

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Citations

Jan 1, 1981·Journal of Immunological Methods·B MicheelH Fiebach
Apr 1, 1980·Journal of Clinical Chemistry and Clinical Biochemistry. Zeitschrift Für Klinische Chemie Und Klinische Biochemie·M Oellerich
Sep 1, 1981·Annals of Clinical Biochemistry·D S SmithJ Landon

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