Chemistry: analytical and immunological testing – Involving an insoluble carrier for immobilizing immunochemicals
Patent
1998-06-01
2000-12-12
Chin, Christopher L.
Chemistry: analytical and immunological testing
Involving an insoluble carrier for immobilizing immunochemicals
436501, 436506, 436507, 436509, 436513, 436528, 436531, 436533, 436534, 436546, 436523, 436805, 436811, 435 5, 435 6, 435 71, 435 794, 435810, G01N 33543, C12Q 170
Patent
active
061597481
ABSTRACT:
Immunoassay methods and apparatus are provided which utilize flow cytometry, coated latex microspheres, and labelled antibodies, to simultaneously detect the presence and amount of several antigens or antibodies in a sample. Microspheres can be sized by forward angle light scatter (FALS) or electronic volume. By combining FALS and fluorescence, it is practical to use beads of several different sizes, colors or shapes, each bead coated with a different protein or antibody, for the simultaneous detection of multiple analytes in a sample. Available auto-sampling systems make it even more appealing in this regard. In accordance with one embodiment, highly purified RNP. Sm, SS-A, SS-B and Scl-70 antigens are bound to 4, 5, 6, 7 and 10 .mu.m latex beads, respectively and stabilized for extended shelflife. Diluted patient serum is placed into test tubes containing a mixture of the five antigen coated beads and incubated. If an antibody is present for a specific antigen, it will bind to that specific bead. After washing the bead/serum mixture to remove residual sample, a second incubation with goat anti-human IgG, conjugated with a fluorochrone, is carried out. This conjugate will bind immunologically to the anti-antigen IgG of the antigen-antibody complex. forming a "sandwich" consisting of bead--antigen--primary antibody--secondary antibody--FITC. The fluorescence intensity is based on the avidity of the bead/antibody/conjugate binding. The samples are analyzed using a flow cytometer.
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AffiniTech, LTD
Chin Christopher L.
Do Pensee T.
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