Process for immobilizing a protein containing substance on a solid phase

• US 5,310,885 A
• Filed: 05/19/1992
• Issued: 05/10/1994
• Est. Priority Date: 12/19/1988
• Status: Expired due to Term

• Current Assignee: Boehringer Mannheim GmbH (Roche Holding AG)
• Original Assignee: Boehringer Mannheim GmbH (Roche Holding AG)
• Inventors: Maier, Josef, Mangold, Dieter, Schlipfenbacher, Reiner
• Primary Examiner(s): Russel, Jeffrey E.
• Application Number:
  US07/887,239
• Time in Patent Office: 721 Days
• Field of Search: 436/528, 436/535, 436/823, 436/530, 530/362, 530/369, 530/403, 530/413, 530/812, 530/813, 530/814, 530/815, 530/817, 530/391.7, 435/175, 435/178, 435/179
• US Class Current: 530/413
• CPC Class Codes: G01N 33/543 with an insoluble carrier for immobilising immunochemicals
• CPC Class Codes: Y10S 530/812 Peptides or proteins is immobilized on, or in, an organic carrier
• CPC Class Codes: Y10S 530/813 Carrier is a saccharide
• CPC Class Codes: Y10S 530/814 Cellulose or derivatives thereof
• CPC Class Codes: Y10S 530/815 Carrier is a synthetic polymer
• CPC Class Codes: Y10S 530/817 Entrapped within the carrier, e.g. gel, hollow fibre

Abstract

The present invention provides a process for immobilizing a protein or protein containing substance. The material to be immobilized is aggregated, contacted in a liquid with a hydrophilic solid phase and the solid phase, after contact has taken place, is dried. The present invention is also concerned with the solid phase prepared by this process and with the use thereof for analytical determination.

Claims

1. Process for immobilizing a protein containing substance onto a solid phase comprising:

(i) forming a water soluble aggregate of a polymer containing more than one molecule of a particular protein;

(ii) contacting a liquid containing said aggregate in dissolved form with a hydrophilic solid phase so as to adsorptively, and non-covalently bind said aggregate to said hydrophilic solid phase; and

(iii) drying said solid phase to form a solid phase having said aggregate immobilized and non-covalently bound thereon.

2. Process of claim 1, wherein said forming of said water soluble aggregate comprises mixing more than one molecule of a particular protein with a polyfunctional linker.

3. Process of claim 1, wherein said drying comprises treating said solid phase to contain less than 7% moisture by weight at 20°

C. and 50% relative humidity.

4. Solid phase having protein bound thereto comprising a hydrophilic solid phase having non-covalently bound thereto a protein aggregate, wherein said aggregate is formed by aggregating more than one molecule of a particular protein to form a polymer containing aggregate prior to contacting said aggregate with said solid phase, wherein said solid phase is dried following contact of said solid phase with said polymer containing aggregate.

5. Solid phase of claim 4, wherein said solid phase is characterized by a moisture content of less than 7% by weight at 20°

C. and 50% relative humidity.

6. Method for determining an analyte in a liquid sample, comprising contacting said liquid sample with the protein aggregate containing solid phase of claim 5, wherein said aggregate comprises a substance which binds with said analyte and determining said analyte bound to said aggregate.

7. Process for immobilizing a protein containing substance onto a solid phase comprising:

(i) forming a water soluble protein homopolymer aggregate;

(ii) contacting a liquid containing said protein homopolymer aggregate in dissolved form with a hydrophilic solid phase so as to adsorptively, and non-covalently bind said protein homopolymer aggregate to said hydrophilic solid phase; and

(iii) drying said solid phase to form a solid phase having said protein homopolymer aggregate immobilized and non-covalently bound thereon.

8. Solid phase having protein bound thereto comprising a hydrophilic solid phase having non-covalently bound thereto a protein homopolymer aggregate, wherein said homopolymer aggregate is formed prior to contacting said homopolymer aggregate to said solid phase, wherein said solid phase is dried following contact of said solid phase with said homopolymer aggregate.

9. Solid phase having protein bound thereto comprising a hydrophilic solid phase having non-covalently bound thereto a protein aggregate, wherein said aggregate is formed by mixing more than one molecule of a particular protein with a polyfunctional linker to form a protein containing aggregate prior to contacting said protein aggregate with said hydrophilic solid phase, wherein said solid phase is dried following contact of said solid phase with said protein aggregate.