Antigen/anti-antigen cleavage

• US 5,429,927 A
• Filed: 11/10/1992
• Issued: 07/04/1995
• Est. Priority Date: 04/09/1990
• Status: Expired due to Term

• Current Assignee: Life Technologies AS (Perspecta, Inc.)
• Original Assignee: Dynel Comapny
• Inventors: Funderud, Steiner, Mortada, Mohamad, Caignault, Laurent, Afseth, John
• Primary Examiner(s): Saunders, David
• Assistant Examiner(s): GRUN, JAMES LESLIE
• Application Number:
  US07/946,330
• Time in Patent Office: 966 Days
• Field of Search: 435/2, 435/7.24, 435/174, 435/176, 435/177, 435/975, 435/965, 435/240.2, 435/239, 435/243, 435/261, 436/526, 436/528, 436/512, 436/518, 530/391.1, 530/863
• US Class Current: 435/7.2
• CPC Class Codes: G01N 33/54326 Magnetic particles
• CPC Class Codes: G01N 33/6857 Antibody fragments
• CPC Class Codes: G01N 33/686 Anti-idiotype
• CPC Class Codes: Y10S 435/975 Kit

Abstract

The present invention provides a method of cleaving an antigen/anti-antigen or hapten/anti-hapten linkage joining two particles comprising reacting said linkage with a secondary antibody, or fragment thereof, binding to said anti-antigen or anti-hapten, and a kit for performing such a method. The method of the invention has particular utility in the separation of cells.

Claims

1. A method of cleaving an antigen/anti-antigen or hapten/anti-hapten linkage joining two particles comprising binding to said anti-antigen or anti-hapten of said linkage a secondary antibody or antibody fragment that specifically binds to one or more sites in said anti-antigen or anti-hapten which are not the antigen-binding or hapten-binding sites and is effective upon binding Said one or more sites to cleave said linkage.

2. A method as claimed in claim 1 wherein the secondary antibody or antibody fragment binds to the Fab region of said anti-antigen or anti-hapten.

3. A method as claimed in claim 1 wherein the secondary antibody and/or anti-hapten or anti-antigen is a complete antibody.

4. A method as claimed in claim 1 wherein the anti-hapten or anti-antigen is an antibody fragment retaining binding activity.

5. A method as claimed in claim 1 wherein the secondary antibody or antibody fragment is an IgG or IgM.

6. A method as claimed in claim 1 wherein the secondary antibody or antibody fragment is polyclonal.

7. A method as claimed in claim 1 wherein in said linkage, the anti-antigen or anti-hapten is attached to a particulate solid support.

8. A method as claimed in claim 1 wherein the linkage joins an anti-antigen or anti-hapten carrying particulate solid support and surface-antigen or hapten carrying cells.

9. A method as claimed in claim 1 wherein the secondary antibody is attached to particles.

10. A method of positively isolating target cells from a mixed population of cells comprising binding an anti-antigen attached to magnetic particles to antigen on said target cells, said anti-antigen being attached to magnetic particles before or after binding to said target cells, thereby binding said particles to said target cells, isolating said magnetic particles and bound target cells from the mixed population of cells by magnetic aggregation, releasing the target cells from said magnetic particles by adding under conditions effective for binding to occur a secondary antibody or antibody fragment that specifically binds to one or more sites in said anti-antigen which are not the antigen-binding sites and that is effective upon binding said one or more sites to cleave the binding between said target cells and said anti-antigen, thereby isolating the released target cells from said mixed population.

11. A method as claimed in claim 10 wherein the secondary antibody or antibody fragment is from a different species from that of the target cell.

12. A kit comprising:

(i) a particulate solid support carrying an anti-antigen or anti-hapten specific for a target antigen or hapten;

(ii) a secondary antibody or antibody fragment that specifically binds to one Or more sites in said anti-antigen or anti-hapten which are not the antigen-binding or hapten-binding sites and is effective upon binding said one or more sites to cleave the antigen/anti-antigen or hapten/anti-hapten linkage formed upon binding of said antigen to Said anti-antigen or Said hapten to said anti-hapten.

13. A kit as claimed in claim 12 wherein the solid support comprises magnetic particles.