Method to prepare magnetic beads conjugated with small compounds

• US 8,901,044 B2
• Filed: 05/11/2010
• Issued: 12/02/2014
• Est. Priority Date: 05/13/2009
• Status: Expired due to Fees

• Current Assignee: Riken
• Original Assignee: Riken
• Inventors: Osada, Hiroyuki, Ohta, Kunihiro, Honda, Kaori, Murayama, Akiho
• Primary Examiner(s): Leavitt, Maria
• Assistant Examiner(s): Bunker, Amy M
• Application Number:
  US12/777,562
• Publication Number:
  US 20100311608A1
• Time in Patent Office: 1,666 Days
• Field of Search: 506/9
• US Class Current: 506/9
• CPC Class Codes: B03C 1/01 by addition of magnetic adjuvants
• CPC Class Codes: B03C 2201/18 Magnetic separation whereby the particles are suspended in a liquid
• CPC Class Codes: G01N 33/54326 Magnetic particles
• CPC Class Codes: G01N 33/54353 with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393

Abstract

It is an object of the present invention to provide a method for stably and efficiently binding a compound to magnetic beads. The present invention relates to a method for producing compound-bound magnetic beads, which comprises: allowing a compound to come into contact with magnetic beads, on the surface of each of which a photoreactive compound has bound; extending the magnetic beads together with the compound on a support; and applying light to the magnetic beads to form a covalent bond between the photoreactive compound and the compound.

Claims

1. A method for producing compound-bound magnetic beads, which comprises:

providing magnetic beads having a photoreactive compound bound on a surface thereof, wherein said photoreactive compound includes;

a compound having an azido group;

a compound having a diazo group or diazirine ring;

a benzophenone, a conjugated ketone, an enone, an aromatic halogen compound or an olefin;

a compound comprising a diazonium group;

or N-(17-amino-3,6,9,12,15-pentaoxaheptadecacil)-4-(3-trifluoromethyl-3H-diazirin-yl)-benzamide;

allowing a low molecular weight compound to come into contact with said magnetic beads;

extending said magnetic beads together with said low molecular weight compound on a support; and

applying light to said magnetic beads and said low molecular weight compound that are extended on said support to form a covalent bond between said photoreactive compound and said low molecular weight compound, wherein a density of said magnetic beads extended on said support upon application of light to said magnetic beads and said low molecular weight compound is not greater than 1×

10⁸ beads/cm².

2. The method according to claim 1, wherein said magnetic beads and said low molecular weight compound are dried, before applying said light thereto.

3. The method according to claim 1, wherein said low molecular weight compound has 0 to 5 amino groups.

4. The method according to claim 3, wherein said low molecular weight compound is a lipid.

5. A kit comprising said magnetic beads, said photoreactive compound, said low molecular weight compound, and an instruction manual, which is used to carry out the method according to claim 1.

7. The method according to claim 1, wherein said low molecular weight compound has 0 amino groups.

6. A method for producing compound-bound magnetic beads comprising:

providing magnetic beads including a photoreactive compound bound on a surface thereof, wherein said photoreactive compound includes;

a compound having an azido group;

a compound having a diazo group or diazirine ring;

a benzophenone, a conjugated ketone, an enone, an aromatic halogen compound or an olefin;

a compound comprising a diazonium group;

or N-(17-amino-3,6,9,12,15-pentaoxaheptadecacil)-4-(3-trifluoromethyl-3H-diazirin-yl)-benzamide;

allowing a lipid compound to come into contact with said magnetic beads, said lipid compound having 0 amino groups;

extending said magnetic beads together with said lipid compound on a support;

applying light to said magnetic beads and said lipid compound that are extended on said support to form a covalent bond between said photoreactive compound and said lipid compound, wherein a density of said magnetic beads extended on said support upon application of light to said magnetic beads and said lipid compound is not greater than 1×

10⁸ beads/cm²;

collecting said magnetic beads; and

subjecting to an antibody library said collected magnetic beads, bonding an antibody from said antibody library to said lipid compound on said magnetic beads and using magnetism to select said magnetic beads having said bound antibody.

8. The method according to claim 6 wherein said step of applying said light occurs while said magnetic beads are extended on said support as a single layer.