Bio-rad Laboratories, Inc.

Bio-red Laboratories, Inc.

Michael I Watkins

Richard B Edwards

Steven R Binder

Renato B Del Rosario

Patrick F Coleman

Suknan S Chang

Patricia A Miranda

Timothy D Knight

Daming Shan

Edward T Liang

Mark A Staples

John M Pocekay

Woei Tan

Clifford R Bartlett

Tony Prestigiacomo

Aleksander Raskin

Gregory A Marr

Xiaoyun Yang

Richard Bruehl

The present invention provides methods and reagents for detecting anti-phospholipid-cofactor protein-antibodies.

Solid phase immobilization of phospholipids and cofactor proteins via covalent attachment

Immunological assays for several biological markers for thyroid disorders in a biological sample are performed in a single test with a combination of sandwich-type, sequential competitive, and serological assays by the use of particles classified into groups that are distinguishable by flow cytometry, one group for the assay of each marker. Each group of particles is coated with a different immunological binding member, and coating densities, co-coating materials, and special buffer solutions are used to adjust for differences in the sensitivities and dynamic ranges of each of the markers in the typical sample.

Multi-analyte diagnostic test for thyroid disorders

Heterogeneous assays for different analytes in a single biological sample are performed simultaneously in a multiplexed assay that combines flow cytometry with the use of magnetic particles as the solid phase and yields an individual result for each analyte. The particles are distinguishable from each other by characteristics that permit them to be differentiated into groups, each group carrying an assay reagent bonded to the particle surface that is distinct from the assay reagents of particles in other groups. The magnetic particles facilitate separation of the solid and liquid phases, permitting the assays to be performed by automated equipment. Assays are also disclosed for the simultaneous detection of antibodies of different classes and a common antigen specificity or of a common class and different antigen specificities. Each type is accomplished by immunological binding at the surfaces of two distinct solid phases in a sequential manner with dissociation of the binding and washing of the solid phase in between the binding steps.

Multiplex flow assays preferably with magnetic particles as solid phase

Water-soluble polymer is added to the liquid phase in a heterogeneous a immunoassay of serum, the polymer having monomers in common with monomers of the solid phase surface. This reduces non-specific binding of IgG's from the serum to the solid phase surface and thereby reduces the occurrence of false positive readings in the immunoassay.

Reducing non-specific binding in immunoassays performed on polymeric solid phases

Multiplex flow assays with magnetic particles as solid phase

Magnetic particles as solid phase for multiplex flow assays

Denaturing agents used to pretreat test samples for immunoassays are neutralized and deactivated by the addition of a treatment agent which is a combination of a buffering agent in acid form and an agent for converting the sulfhydryl groups of the denaturing agents to a non-active form such as disulfides or alkylthio groups. The treatment agent serves this function without having sufficient denaturing activity by itself to interfere with subsequent steps of the assay procedure. Also disclosed is a protocol for a vitamin B.sub.12 assay which involves a sequential rather than competitive binding of the B.sub.12 with excess intrinsic factor followed by the binding of excess enzyme-labeled B.sub.12 to any remaining intrinsic and immobilizing such intrinsic factor on a solid phase. Various problems associated with B.sub.12 assays are avoided by this technique.

B.sub.12 enzyme imunoassay and sample pretreatment

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Vacaville, CA, United States of America

Michael I Watkins