Method and apparatus for protein manipulation
AssigneeLarge Scale Proteomics Corporation
InventorN. Leigh Anderson
Life of this patent
Estimating the term…
Abstract
A method and apparatus for extracting, identifying, and manipulating proteins or peptides from a solution uses paramagnetic beads having a coating with an affinity for the target component. In one embodiment, paramagnetic beads coated with C18 are used to adsorb proteins and peptides. The beads can be used to purify, immobilize and assay antibodies. By cycling the beads, many times greater molar amount of binding partner may be separated from a solution. A magnetic probe is used to capture the beads and transfer the beads to selected processing stages.
Claims46
A method of recovering a target component from a sample, said method comprising the steps of:
- providing a solution containing said sample;
- contacting said solution with an amount of magnetically responsive beads, said beads having an outer surface with a polymeric coating having pendant hydrophobic groups, said hydrophobic groups having a binding affinity for said target component, said beads being contacted with said solution for sufficient time to bind said target component to said beads;
- and
- recovering said beads from said solution.
A method of recovering a protein or peptide from a sample, said method comprising the steps of:
- providing a solution containing said sample;
- contacting said solution with an amount of magnetically responsive beads, said beads having an outer surface with a polymeric coating having a binding partner with a binding affinity for said protein or peptide, said beads being contacted with said solution for sufficient time and under conditions for said binding partner to bind said protein or peptide to said beads, wherein said coating or said binding partner on said beads is reactive with said protein or peptide and forms a covalent chemical bond with said protein or peptide;
- separating said beads from said solution;
- and
- cleaving said proteins or peptides from said beads.
A method of recovering a plurality of target components from a sample, said method comprising the steps of:
- providing a solution of said sample containing first and second target components;
- contacting said solution with an amount of first magnetically responsive beads comprising a coating with a specific binding affinity for a first target component, and an amount of second magnetically responsive beads having a coating with a specific binding affinity for a second target component,
- simultaneously contacting said beads with said solution for sufficient time to bind said first and second target components to said first and second magnetically responsive beads, respectively;
- and
- recovering said beads from said solution.
A method of recovering a target component from a sample, said method comprising the steps of:
- providing a solution and a support medium in said solution, said support medium containing said sample;
- contacting said solution with an amount of magnetically responsive beads, said beads having an outer surface with a polymeric coating having a binding affinity for said target component,
- applying an electric current to said solution to cause said target component to diffuse from said support medium into said solution, said beads being contacted with said solution for sufficient time to bind said target component to said beads;
- and
- recovering said beads from said solution.
A method of recovering a target component comprising the steps of:
- contacting a solution or dispersion of a target component with an amount of magnetically responsive beads having a coating with a binding affinity for said target component, said contacting step being for sufficient time for said target component to bind to said beads;
- aspirating said solution or dispersion containing said beads through a tube having a filter device and collecting said beads on said filter device;
- washing said beads inside said tube by passing a wash solution through said tube, and
- eluting said target component from said beads.
A method of continuously recovering a target component from a liquid sample comprising the steps of:
- providing a liquid sample containing said target component in a sample vessel and contacting said liquid sample with an effective amount of magnetically responsive beads having a coating with a binding affinity for said target component, said contacting step being for sufficient time to enable said target component to bind to said coating on said beads;
- transferring said beads to an eluting vessel containing a liquid eluting agent and eluting said target component from said beads;
- and
- returning said beads to said sample vessel containing said liquid sample.
A method of recovering a denatured protein from a sample, said method comprising the steps of:
- providing a solution containing said denatured protein from said sample;
- contacting said solution with an amount of magnetically responsive beads, said beads having an outer surface having a specific binding affinity for native protein, said beads being contacted with said solution for sufficient time to bind said denatured protein to said beads;
- recovering said beads from said solution, and
- eluting said denatured protein from said beads.
Dependencies are inferred from the claim text.