Boehringer Mannheim Gmbh

Michael Berger

Arno Deger

Josef Maier

Dittmar Schlieper

Beatus Ofenloch-Hahnle

Fran&ccedil;ois Guillot

The invention concerns a homogeneous immunoassay based on agglutination using an Fab&prime;-biotin material and agglutinatable particles which carry streptavidin or avidin. The invention also concerns the Fab&prime;-biotin which is bound or linked via linkage groups to a label compound which can electrochemiluminescence and the particles having avidin or streptavidin on their surface are magnetic.

Method for carrying out a homogeneous-immunoassay based on agglutination

A specific binding pair is bound to an insoluble carrier for use in determining an analyte such as in an immunoassay. The carrier is coated with a first polymer containing a protein polymer having a molecular weight of at least about 20,000 and molecules of a first member of a specific binding pair. A second polymer containing a second member of the specific binding pair is bound to the first member on the carrier by binding of the first and second members of the specific binding pair. The first polymer is preferably more hydrophobic than the second polymer. The protein polymer can be prepared by cross-linking hydrophobic protein molecules of 10,000 to 700,000 molecular weight with a bifunctional or polyfunctional compound to obtain a protein polymer of 200,000 to 20,000,000 molecular weight. The second polymer can be the second member of the specific binding pair or the second member cross-linked with a linker or the second member cross-linked to a hydrophobic protein.

Preparation of a solid phase matrix containing a bound specific binding

The present invention provides a process for the detection of analytes in body fluids containing free biotin by immunoassay with the use of biotin conjugates for the labelling, immobilization or complexing of immunological reagents, wherein the biotin present in free form is removed from the immunological reaction by incubating the sample solution with polymer particles consisting of a core and a covering which, as core, contain a polymer which has a plurality of binding sites for biotin and, as covering, at least one layer of protein, peptide, carbohydrate or co-polymer of carbohydrate and amino acids, the layer thickness of the covering being so adjusted that the coating is permeable for the free biotin but not for the biotin conjugate.

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Penzberg, Germany

Michael Berger