Method of performing affinity separation using immobilized flocculating agent on chromatographic support
First Claim
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1. A method of performing an affinity separation comprising the steps of;
- a. forming a primary chromatographic support with a flocculating agent immobilized thereon, the primary support selected from the group consisting of particulate resins and porous plastic materials;
b. forming a secondary support consisting of microparticles having ligand or binder for the ligand affixed thereon;
c. contacting the primary support of step a with the secondary support of step b to form an affinity support with said microparticles flocculated onto said primary support; and
d. contacting a sample suspected of containing a binder or ligand complementary to the ligand or binder attached on the affinity support of step c to separate said binder or ligand from the sample.
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Abstract
Disclosed is a primary chromatographic support containing an immobilized flocculating agent for use as a separation media for a secondary support consisting of microparticles used in affinity separation and heterogeneous immunoassays. In a specific embodiment, a flocculating agent such as polyethyleneimine is immobilized on a chromatographic resin packed in a column. The column so formed is then used to trap microparticles having an affinity ligand or binder for the ligand affixed thereon. Such microparticles are used as a solid support for the affinity reaction involved in a variety of immunoassay formats.
137 Citations
8 Claims
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1. A method of performing an affinity separation comprising the steps of;
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a. forming a primary chromatographic support with a flocculating agent immobilized thereon, the primary support selected from the group consisting of particulate resins and porous plastic materials; b. forming a secondary support consisting of microparticles having ligand or binder for the ligand affixed thereon; c. contacting the primary support of step a with the secondary support of step b to form an affinity support with said microparticles flocculated onto said primary support; and d. contacting a sample suspected of containing a binder or ligand complementary to the ligand or binder attached on the affinity support of step c to separate said binder or ligand from the sample. - View Dependent Claims (2, 3, 4)
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5. A method of performing affinity separation comprising the steps of:
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a. forming a primary chromatographic support with a flocculating agent immobilized thereon, the primary support selected from the group consisting of particulate resins and porous plastic materials; b. forming a secondary support consisting of microparticles having ligand or binder for the ligand affixed thereon; c. contacting a sample suspected of containing a binder or ligand complementary to the ligand or binder attached on the microparticles of step b with said microparticles to capture the binder or ligand from the sample onto the microparticles; and d. contacting the reaction .mixture of step c with the primary support of step a to trap the microparticles. - View Dependent Claims (6, 7, 8)
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Specification