Abstract: A new surface immunoassay method is performed with two test surfaces. The method is used where serum samples may contain both an antibody for which it is desirable to test, and also competing sera constituents which may be present in indeterminate amounts. In accordance with the method, two different test surfaces are contacted with the same serum sample, and preferably the same aliquot of the serum. The first surface is treated to bind a broad spectrum of serum components including both the component to be tested for and also the competing components. The second surface is designed to bind the competing components but no substantial amount of the serum component to be tested for. The immunoassay is conducted with a labeled antibody, and quantitative measurement of the immunoassay is obtained by detecting the quantitative difference in the amount of labeled antibody on the two surfaces which have been subjected to the same serum sample.

Abstract: An assay method suitable for antigens (or haptens) including proteins and polypeptides utilizing antibodies for the unknown protein or polypeptide in which the antibodies are covalently bound to water insoluble hydrophilic polymeric particles. Appropriate fluorescently labeled immune reactants are introduced during the method and together with the unknown antigen are immunologically bound, directly or indirectly, to the particles for separation from the reaction liquid. The particles have a size of about 0.1-10 microns and can be resuspended for direct measurement of the fluorescent labels in a fluorometer from which the unknown can be determined.

Abstract: An assay method suitable for antigens (or haptens) including proteins and polypeptides utilizing antibodies for the unknown protein or polypeptide in which the antibodies are covalently bound to water insoluble hydrophylic polymeric particles. Appropriate fluorescently labeled immune reactants are introduced during the method and together with the unknown antigen are immunologically bound, directly or indirectly, to the particles for separation from the reaction liquid. The particles have a size of about 0.1-10 microns and can be resuspended for direct measurement of the fluorescent labels in a fluorometer from which the unknown can be determined.

Abstract: This invention provides a method for the radioimmunoassay of thyroid-stimulating hormone which utilizes a rapid and convenient version of a double antibody procedure. Highly purified second antibody is bound, by means of covalent bonds, to hydrolyzed polyacrylamide particles to produce a two-phase system. The solid phase comprises immobilized second antibody bound to the reaction product of labeled and unlabeled thyroid-stimulating hormone with the first antibody (first antibody-antigen complex) and the liquid phase comprises free (unbound) labeled and unlabeled thyroid-stimulating hormone. The two phases are separated and the radioactivity of either phase is measured.