Method of coupling a protein to an epoxylated latex

• US 4,210,723 A
• Filed: 11/25/1977
• Issued: 07/01/1980
• Est. Priority Date: 07/23/1976
• Status: Expired due to Term

• Current Assignee: The Dow Chemical Company (Dow, Inc.)
• Original Assignee: The Dow Chemical Company (Dow, Inc.)
• Inventors: Dorman, Linneaus C., Mani, Inder
• Primary Examiner(s): Naff, David M.
• Application Number:
  US05/854,930
• Time in Patent Office: 949 Days
• Field of Search: 195/63, 195/68, 195/DIG. 11, 195/103.5 A, 195/99, 260/112 R, 424/12, 23/230 B, 23/915
• US Class Current: 435/180
• CPC Class Codes: C12N 11/082 obtained by reactions only involving carbon-to-carbon unsaturated bonds
• CPC Class Codes: C12N 11/087 Acrylic polymers
• CPC Class Codes: G01N 33/76 Human chorionic gonadotropin including luteinising hormone, follicle stimulating hormone, thyroid stimulating hormone or their receptors
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/814 Pregnancy
• CPC Class Codes: Y10S 436/817 Steroids or hormones
• CPC Class Codes: Y10S 436/818 Human chorionic gonadotropin

Abstract

Proteins are coupled to polymer particles containing free epoxy groups in a latex. The polymer particles are about 0.15 to 1.5 micrometers in diameter, and have an inner core and an outer shell which contains the free epoxy groups. The coupled proteins are particularly useful in carrying out immunological tests.

Claims

1. A method for coupling a protein having a reactive group containing a labile hydrogen to polymer particles containing free epoxy groups in a latex wherein the polymer particles in the latex range in size from about 0.15 to 1.5 micrometers in diameter and have an inner core and an outer shell containing free epoxy groups, said inner core being formed by the polymerization or copolymerization of one or more hard monomers and optionally soft monomer to produce a latex containing polymer particles that are said inner core and said outer shell being formed by copolymerization in the presence of said inner core polymer particles one or more hard monomers and a copolymerizable ethylenically unsaturated compound having free epoxy groups and optionally soft monomer, said method comprising reacting said polymer particles containing free epoxy groups in the latex and said protein at a pH of 7.0-9.0 for a time sufficient to form a covalent bond between the reactive group of the protein and the free epoxy group of the polymer particles and removing the unreacted protein from the latex-protein conjugate.

2. The method of claim 1 wherein the latex containing the polymer particles having free epoxy groups is a monodispersed latex and the reactive group on the protein is amino.

3. The method of claim 1 wherein the outer shell on the polymer particles present in the latex is styrene-glycidyl methacrylate copolymer.

4. The method of claim 3 wherein the protein is human chorionic ganadotropin.

5. The method of claim 3 wherein the protein is insulin.

6. The method of claim 3 wherein the protein is an enzyme.

7. The method of claim 4 wherein the coupling site density of the styrene-glycidyl methacrylate copolymer outer shell is about one epoxy unit per 5 to 40 square angstrom units of particle surface area and the amount of human chorionic gonadotropin used to prepare the polymer particle protein conjugate is 1,000 to 25,000 international units per gram of polymer particles.

8. A protein polymer particle conjugate in a latex formed by the method of claim 2.

9. The protein-latex conjugate of claim 8 wherein the protein is human chorionic gonadotropin.

10. The protein-polymer particle conjugate in a latex of claim 8 wherein the protein is insulin and the latex contains polymer particles having an outer shell of styrene-glycidyl methacrylate copolymer.

11. The protein polymer particle conjugate in a latex of claim 8 wherein the latex is dyed.

12. A kit for immunologically testing for pregnancy which comprises as a first reagent the protein-polymer particle conjugate in a latex of claim 9 and as a second reagent, anti-human chorionic gonadotropin serum.