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4959310 Monoclonal antibodies to soybean Kunitz trypsin inhibitor and immunoassay methods
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New Patents
A method and device for determining the presence of an analyte in a sample suspected of containing the analyte are disclosed. The method involves contacting a test solution containing the sample, an antibody for the analyte, and a conjugate of the analyte and a label with a contact portion of a strip of bibulous material capable of being traversed by the test solution through capillary action. The strip contains a first receptor capable of binding to the conjugate. The first receptor is non-diffusively bound to a situs on the strip separate from the contact portion of the strip. The strip further contains a second receptor capable of binding the antibody to the analyte between the situs and the contact portion. The second receptor is non-diffusively bound to the strip. At least a portion of the test solution is allowed to traverse the strip by capillary action and thereby contact the situs. The strip is exposed to a signal producing means capable of interacting with the label to produce a signal in relation to the amount of analyte in the test solution. The signal produced at the situs is then detected.
4959308 IMMUNOASSAY FOR ANTIBODIES BINDING PLATELETS Daryl M Ogden assigned to Board of Regents The University of Texas System The present invention involves a method for producing a substrate useful in a system for the detection of antibodies directed against platelet antigens. This method comprises several steps. A platelet sample of interest is initially treated with an aqueous solution comprising a dialyzable nonionic detergent. This initial treatment is under conditions to solubilize platelet components and produce a platelet lysate. Such conditions may involve treatment of a platelet sample with an aqueous solution comprising nonionic detergent at a concentration between about 0.2% and about 0.5%. Platelet antigens are most preferably solubilized for about 30 min and at about 0 degrees C. in an aqueous solution comprising about I mg dialyzable nonionic detergent per mg platelet protein. The partially purified platelet antigens resulting from these manipulations are then preferably affixed to a solid matrix. The solid matrix preferably comprises nitrocellulose
paper, polystyrene or latex but may be any solid matrix suitable for the abstraction from a biological sample and/or assay of antibodies binding to the affixed purified platelet antigens.
4959310 MONOCLONAL ANTIBODIES TO SOYBEAN KUNITZ TRYPSIN INHIBITOR AND IMMUNOASSAY METHODS David Brandon, Anne H Bates, Mende Friedman assigned to The United States of America as represented by the Secretary of Agriculture Hybrid cell lines (hybridomas) which produce and secrete monoclonal antibodies with three distinct patterns of recognition are described. The first specificity pattern is defined by antibodies which (1) recognize Kunitz trypsin inhibitor (KTI), one of the principal protease inhibitors found in soybeans, but do not detect the Bowman-Birk inhibitors (BB1), the other major class of protease inhibitors in soybeans; (2) bind to native KTI isoforms a and b but do not react with KTI isoforms a and b which have been denatured by moist heat or alkaline treatment or which have been subjected to disulfide exchange; and (3) do not recognize native KTI isoform c. The second specificity pattern is defined by antibodies that (1) recognize KTI, but do not bind BBI; (2) bind native KTI isoforms a and c, but do not bind strongly to KTI isoforms a and c which have been denatured by moist heat or alkaline treatment or which have been subjected to disulfide exchange; (3) do not bind KT1 isoform b; and (4) bind only weakly to KTI when KT1 is complexed with trypsin or a similar enzyme. The third specificity pattern is defined by antibodies that (1) recognize KTI, but do not bind BBI; (2) bind equivalently to native KT1 isoforms a, b, and c, but do not bind to KTI which has been denatured by moist heat or alkaline treatment or which has been subjected to disulfide exchange; and (3) bind equivalently to uncomplexed KTI and KTI complexed with trypsin or a similar enzyme. Immunoassay methods using the monoclonal antibodies to analyze native KTI specifically in soy-derived foodstuffs and in tissues of soybean plants, to determine isoform content of a sample, and to determine the amount of KT1 complexed with trypsin in a sample are described.
New Patents
A method and device for determining the presence of an analyte in a sample suspected of containing the analyte are disclosed. The method involves contacting a test solution containing the sample, an antibody for the analyte, and a conjugate of the analyte and a label with a contact portion of a strip of bibulous material capable of being traversed by the test solution through capillary action. The strip contains a first receptor capable of binding to the conjugate. The first receptor is non-diffusively bound to a situs on the strip separate from the contact portion of the strip. The strip further contains a second receptor capable of binding the antibody to the analyte between the situs and the contact portion. The second receptor is non-diffusively bound to the strip. At least a portion of the test solution is allowed to traverse the strip by capillary action and thereby contact the situs. The strip is exposed to a signal producing means capable of interacting with the label to produce a signal in relation to the amount of analyte in the test solution. The signal produced at the situs is then detected.
4959308 IMMUNOASSAY FOR ANTIBODIES BINDING PLATELETS Daryl M Ogden assigned to Board of Regents The University of Texas System The present invention involves a method for producing a substrate useful in a system for the detection of antibodies directed against platelet antigens. This method comprises several steps. A platelet sample of interest is initially treated with an aqueous solution comprising a dialyzable nonionic detergent. This initial treatment is under conditions to solubilize platelet components and produce a platelet lysate. Such conditions may involve treatment of a platelet sample with an aqueous solution comprising nonionic detergent at a concentration between about 0.2% and about 0.5%. Platelet antigens are most preferably solubilized for about 30 min and at about 0 degrees C. in an aqueous solution comprising about I mg dialyzable nonionic detergent per mg platelet protein. The partially purified platelet antigens resulting from these manipulations are then preferably affixed to a solid matrix. The solid matrix preferably comprises nitrocellulose
paper, polystyrene or latex but may be any solid matrix suitable for the abstraction from a biological sample and/or assay of antibodies binding to the affixed purified platelet antigens.
4959310 MONOCLONAL ANTIBODIES TO SOYBEAN KUNITZ TRYPSIN INHIBITOR AND IMMUNOASSAY METHODS David Brandon, Anne H Bates, Mende Friedman assigned to The United States of America as represented by the Secretary of Agriculture Hybrid cell lines (hybridomas) which produce and secrete monoclonal antibodies with three distinct patterns of recognition are described. The first specificity pattern is defined by antibodies which (1) recognize Kunitz trypsin inhibitor (KTI), one of the principal protease inhibitors found in soybeans, but do not detect the Bowman-Birk inhibitors (BB1), the other major class of protease inhibitors in soybeans; (2) bind to native KTI isoforms a and b but do not react with KTI isoforms a and b which have been denatured by moist heat or alkaline treatment or which have been subjected to disulfide exchange; and (3) do not recognize native KTI isoform c. The second specificity pattern is defined by antibodies that (1) recognize KTI, but do not bind BBI; (2) bind native KTI isoforms a and c, but do not bind strongly to KTI isoforms a and c which have been denatured by moist heat or alkaline treatment or which have been subjected to disulfide exchange; (3) do not bind KT1 isoform b; and (4) bind only weakly to KTI when KT1 is complexed with trypsin or a similar enzyme. The third specificity pattern is defined by antibodies that (1) recognize KTI, but do not bind BBI; (2) bind equivalently to native KT1 isoforms a, b, and c, but do not bind to KTI which has been denatured by moist heat or alkaline treatment or which has been subjected to disulfide exchange; and (3) bind equivalently to uncomplexed KTI and KTI complexed with trypsin or a similar enzyme. Immunoassay methods using the monoclonal antibodies to analyze native KTI specifically in soy-derived foodstuffs and in tissues of soybean plants, to determine isoform content of a sample, and to determine the amount of KT1 complexed with trypsin in a sample are described.