초록 ▼

Methods of determining the ratio of apolipoprotein B-100 to apolipoprotein A-I using ELISA techniques in conjunction with monoclonal paratopic molecules are disclosed as are diagnostic systems useful in performing those determinations. Monoclonal paratopic molecules secreted by hybridomas having ATC

Methods of determining the ratio of apolipoprotein B-100 to apolipoprotein A-I using ELISA techniques in conjunction with monoclonal paratopic molecules are disclosed as are diagnostic systems useful in performing those determinations. Monoclonal paratopic molecules secreted by hybridomas having ATCC accession numbers HB 8742, HB 8746, HB 9200 and HB 9201 are utilized.

대표청구항 ▼

In a method for assaying for a marker of abnormal lipid metabolism comprising the steps of assaying for the amount of human apolipoprotein B-100 and apolipoprotein A-I per unit volume of a blood sample and determining the ratio of apolipoprotein B-100 to apolipoprotein A-I in the sample, the improve

In a method for assaying for a marker of abnormal lipid metabolism comprising the steps of assaying for the amount of human apolipoprotein B-100 and apolipoprotein A-I per unit volume of a blood sample and determining the ratio of apolipoprotein B-100 to apolipoprotein A-I in the sample, the improvement that comprises: (a) assaying a first aliquot of an apolipoprotein B-100-containing liquid blood sample from a person for the amount of apolipoprotein B-100 present by: (i) admixing said first liquid sample aliquot with a solid support consisting essentially of a solid matrix having solid phase-bound first monoclonal paratopic molecules that immunoreact with apolipoprotein B-100 and are secreted by one of the hybridomas having ATCC accession numbers HB 8742 or HB 8746 to form a first solid-liquid phase admixture, the surface of said support having blocked non-specific binding sites; (ii) maintaining said first solid-liquid phase admixture under biological assay conditions for a predetermined period of time sufficient for said first paratopic molecules to immunoreact with apolipoprotein B-100 present in the sample aliquot and form a solid phase-bound immunoreactant that contains substantially all apolipoprotein B-100 present in said sample aliquot; (iii) admixing apolipoprotein B-100 in said first liquid sample aliquot with liquid phase second monoclonal paratopic molecules that immunoreact with apolipoprotein B-100, are secreted by one of the hybridomas having ATCC accession numbers HB 8742 or HB 8746 but are not utilized in step (a)(i), and are operatively linked to an enzyme indicating means to form a second admixture; (iv) maintaining said second admixture under biological assay conditions for a predetermined period of time sufficient for said second indicating means-linked paratopic molecules to form an immunoreactant that contains substantially all apolipoprotein B-100 in the sample aliquot; (v) separating the solid and liquid phses that result from above steps (a)(i-iv); and (vi) determining the amount of indicating means-linked apolipoprotein B-100-containing immunoreactant present in the separated solid phase, and thereby the amount of apolipoprotein B-100 in a unit volume of sample; (b) assaying a second aliquot of said liquid blood sample that contains apolipoprotein A-I and is free from unmasking treatment for the amount of apolipoprotein A-I present by: (i) admixing said second liquid sample aliquot with a solid support consisting essentially of a solid matrix having solid phase-bound third monoclonal paratopic molecules that immunoreact with apolipoprotein A-I and are secreted by one of the hybridomas having ATCC accession numbers HB 9200 or HB 9201 to form a third, solid-liquid phase, admixture, the surface of said solid support having blocked non-specific binding sites; (ii) maintaining said third, solid-liquid phase, admixture under biological assay conditions for a predetermined period of time sufficient for said third paratopic molecules to immunoreact with apolipoprotein A-I present in the aliquot sample and form a solid phase-bound immunoreactant that contains substantially all apolipoprotein A-I in the sample aliquot; (iii) admixing apolipoprotein A-I in said second liquid sample aliquot with liquid phase fourth monoclonal paratopic molecules that immunoreact with apolipoprotein A-I, are secreted by one of the hybridomas having ATCC accession numbers HB 9200 or HB 9201 but are not utilized in step (b)(i), and are operatively linked to an enzyme indicating means to form a fourth admixture; (iv) maintaining said fourth admixture under biological assay conditions for a predetermined period of time sufficient for said fourth, indicating means-linked paratopic molecules to form an immunoreactant with substantially all apolipoprotein A-I present in the sample aliquot; (v) separating the solid and liquid phases that result from steps (b)(i-iv); and (vi) determining the amount of indicating means-linked apolipoprotein A-I-containing immunoreactant present in the separated solid phase, and thereby the amount of apolipoprotein A-I in a volume of sample.