Substrate and method of immobilizing protein

• US 20070055013A1
• Filed: 02/21/2006
• Published: 03/08/2007
• Est. Priority Date: 02/21/2005
• Status: Abandoned Application

• Current Assignee: Kyushu University, Nippon Sheet Glass Company Limited
• Original Assignee: Kyushu University, Nippon Sheet Glass Company Limited
• Inventors: Nakamura, Koichiro, Fujita, Koji, Doi, Satoshi, Goto, Masahiro, Kamiya, Noriho, Tanamura, Koki, Kawata, Noriyuki
• Application Number:
  US11/358,438
• Publication Number:
  US 20070055013A1
• Time in Patent Office: Days
• US Class Current: 525/54.100
• CPC Class Codes: G01N 33/54353 with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393

Abstract

A substrate for the immobilization of proteins is made by treating a carrier with an amino group-containing silicon compound represented by the following general formula:
(RO)₃Si—(CH₂)_k—(C₆H₄)₁—(CH₂)_m—(NHCH₂CH₂)_n—NH₂ (wherein R is an alkyl group, and k is 1, 2, 3 . . . , l is 0, 1, 2, 3 . . . , m is 0, 1, 2, 3 . . . , and n is 1, 2, 3 . . . ).

Claims

1. A substrate having amino groups on a surface thereof, said substrate being prepared by treating a carrier with an amino group-containing silicon compound represented by the following general formula:

(RO)₃Si—

(CH₂)_k—

(C₆H₄)₁—

(CH₂)_m—

(NHCH₂CH₂)_n—

NH₂

to introduce the amino groups onto a surface of the carrier, wherein R is an alkyl group, k is an integer ≧

1, l is an integer ≧

0, m is an integer ≧

0, and n is an integer ≧

1.

2. The substrate according to claim 1, wherein n is 3-20.

3. The substrate according to claim 1, wherein the amino group-containing silicon compound is (3-trimethoxysilylpropyl)-diethylene triamine.

4. A method of immobilizing a protein on a substrate having amino groups on a surface thereof, the method comprising contacting the substrate with a buffer solution comprising the protein, wherein the substrate is made by treating a carrier with an amino group-containing silicon compound represented by the following general formula:

(RO)₃Si—

(CH₂)_k—

(C₆H₄)₁—

(CH₂)_m—

(NHCH₂CH₂)_n—

NH₂

to introduce amino groups onto a surface of the carrier, wherein R is an alkyl group, k is an integer ≧

1, l is an integer ≧

0, m is an integer ≧

0, and n is an integer ≧

1.

5. The method according to claim 4, wherein the buffer solution comprises a protein, a biopolymer or a biological sample.

6. The method according to claim 4, wherein the buffer solution comprises a biological sample derived from is at least one source selected from the group consisting of cells, microorganisms, tissues, and organs.

7. The method according to claim 4, wherein when l in the general formula is more than 1, the immobilization comprises a hydrophobic phenyl group interaction.

8. The method according to claim 4, wherein a pH of the protein-containing buffer solution satisfies a relation of pKa>

pH>

pI, wherein pI is an isoelectric point of the protein and pKa is an acid dissociation constant of the amino group.

9. The method according to claim 4, wherein the immobilization comprises electrostatic immobilization.

10. The method of claim 4, wherein said contacting comprises immersing the substrate in the buffer solution.