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US20020028486A1 - Antibodies having modified carbohydrate content and methods of preparation and use - Google Patents

Antibodies having modified carbohydrate content and methods of preparation and use
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US20020028486A1
US20020028486A1US09/835,461US83546101AUS2002028486A1US 20020028486 A1US20020028486 A1US 20020028486A1US 83546101 AUS83546101 AUS 83546101AUS 2002028486 A1US2002028486 A1US 2002028486A1
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Prior art keywords
antibody
carbohydrate
recognition site
carbohydrate recognition
substance
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Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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US09/835,461
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Sherie Morrison
Vernon Oi
Paul Hinton
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Columbia University in the City of New York
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Columbia University in the City of New York
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Priority to US09/835,461priorityCriticalpatent/US20020028486A1/en
Publication of US20020028486A1publicationCriticalpatent/US20020028486A1/en
Assigned to NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENTreassignmentNATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENTEXECUTIVE ORDER 9424, CONFIRMATORY LICENSEAssignors: COLUMBIA UNIVERSITY NEW YORK MORNINGSIDE
Assigned to NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENTreassignmentNATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENTCONFIRMATORY LICENSE (SEE DOCUMENT FOR DETAILS).Assignors: COLUMBIA UNIVERSITY NEW YORK
Abandonedlegal-statusCriticalCurrent

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Abstract

This invention provides a method of altering the affinity of an antibody for the antigen to which it is directed which comprises introducing into the variable region of the antibody a carbohydrate recognition site under conditions such that a carbohydrate binds to the site and thus attaches to the antibody. This invention also provides a method of modifying the carbohydrate content of an antibody which comprises deleting from a constant region of the antibody a carbohydrate recognition site which naturally occurs in such constant region of such antibody. Antibodies, e.g., monoclonal antibodies and human monoclonal antibodies, diagnostic test kits, DNA encoding antibodies, therapeutic agents, and methods for detecting the presence of a substance in a sample, and for recovering and purifying a substance from a sample are also provided.

Description

Claims (49)

What is claimed is:
1. A method of altering the affinity of an antibody for the antigen to which it is directed which comprises introducing into the variable region of the antibody a carbohydrate recognition site under conditions such that a carbohydrate binds to the site and thus attaches to the antibody.
2. A method ofclaim 1, wherein the antibody is a monoclonal antibody
3. A method ofclaim 1, wherein the antibody is a human antibody.
4. A method ofclaim 1, wherein the carbohydrate recognition site is introduced into the second hypervariable region of the antibody.
5. A method ofclaim 1, wherein the carbohydrate recognition site is introduced into a variable region of the antibody which does not normally include a carbohydrate recognition site.
6. A method ofclaim 1, wherein the carbohydrate recognition site is introduced into the variable region of the antibody by site-directed mutagenesis of DNA encoding the variable region of the antibody.
7. A method of producing an antibody which may be more readily recovered or purified which comprises introducing into the variable region of the antibody a carbohydrate recognition site under conditions such that a carbohydrate binds to the site and thus attaches to the antibody.
8. A method ofclaim 7, wherein the antibody is a monoclonal antibody.
9. A method ofclaim 7, wherein the antibody is a human antibody.
10. A method ofclaim 7, wherein the carbohydrate recognition site is introduced into the second hypervariable region of the antibody.
11. A method ofclaim 7, wherein the carbohydrate recognition site is introduced into a variable region of the antibody which does not normally include a carbohydrate recognition site.
12. A method ofclaim 5, wherein the carbohydrate recognition site is introduced into the variable region of the antibody by site directed mutagenesis of DNA encoding the variable region of the antibody.
13. An antibody which does not occur in nature and which comprises a carbohydrate recognition site genetically engineered into a variable region of the antibody which does not naturally include a carbohydrate recognition site in such variable region.
14. A monoclonal antibody ofclaim 13.
15. A human antibody ofclaim 13.
16. An antibody ofclaim 13, characterized by enhanced affinity for the antigen to which it is directed.
17. An antibody ofclaim 13 labeled with a detectable marker.
18. An antibody ofclaim 17, wherein the detectable marker is a chromophore, fluorophore or radioactive moiety.
19. A solid support to which an antibody ofclaim 16 is bound.
20. An altered antibody prepared by the method ofclaim 1.
21. An antibody ofclaim 13 further comprising carbohydrate attached to the carbohydrate recognition site genetically engineered into the variable region of the antibody.
22. An antibody ofclaim 13, wherein the variable region is the second hypervariable region of the antibody.
23. DNA encoding the antibody ofclaim 21.
24. A method of modifying the carbohydrate content of an antibody which comprises deleting from a constant region of the antibody a carbohydrate recognition site which naturally occurs in such constant region of such antibody.
25. A method ofclaim 24, wherein the antibody is a monoclonal antibody.
26. A method ofclaim 24, wherein the antibody is a human antibody.
27. A method ofclaim 24, wherein the deletion of the carbohydrate recognition site from the constant region of the antibody is effected by site-directed mutagenesis of DNA encoding such constant region of the antibody.
28. A method of modifying the carbohydrate content of an antibody which comprises adding to a constant region of the antibody a carbohydrate recognition site which does not naturally occur in such constant region of such antibody.
29. A method ofclaim 28, wherein the addition of the carbohydrate recognition site to the antibody is effected by site-directed mutagenesis of DNA encoding such constant region of the antibody.
30. A method of modifying the biological effector function of an antibody which comprises modifying the carbohydrate content of the antibody using the method ofclaim 24.
31. A human antibody which does not occur in nature and which is characterized by the absence of a carbohydrate recognition site in a constant region of the antibody which naturally includes such a carbohydrate recognition site in such constant region.
32. A human monoclonal antibody ofclaim 31.
33. A human antibody which does not occur in nature and which is characterized by the presence of a carbohydrate recognition site in a constant region of the antibody which does not naturally include such carbohydrate recognition site in such constant region.
34. A human monoclonal antibody ofclaim 33.
35. A human antibody ofclaim 33 having carbohydrate attached to such carbohydrate recognition site.
36. A labeled antibody ofclaim 35, wherein the label is bound to such carbohydrate.
37. A therapeutic agent comprising the antibody ofclaim 35 and a therapeutic ligand, the ligand being bound to such carbohydrate attached to such antibody.
38. A modified antibody prepared by the method ofclaim 24.
39. A modified antibody prepared by the method ofclaim 28.
40. DNA encoding the antibody ofclaim 31.
41. DNA encoding the antibody ofclaim 33.
42. A more sensitive method for detecting the presence of a substance in a sample which comprises contacting the sample with an antibody ofclaim 17 directed to the substance, under conditions such that substance in the sample forms a detectable complex with the antibody, and detecting the complex and thus the substance.
43. A method ofclaim 42, wherein the antibody is bound to a solid support.
44. A quantitative method ofclaim 42, wherein the amount of complex formed is determined and thereby the amount of the substance present in the sample.
45. A method of recovering a substance from a sample containing the substance which comprises contacting the sample with an antibody ofclaim 13 directed to the substance under conditions such that substance in the sample forms a complex with the antibody and recovering the substance from the resulting complex.
46. A chromatographic method ofclaim 45, wherein the antibody is bound to a solid support.
47. A purification method for a substance which comprises recovering the substance using the method ofclaim 45 from a sample under conditions such that the substance is recovered in purified form.
48. A diagnostic test kit comprising the antibody ofclaim 13.
49. A diagnostic test kit comprising the antibody of claim17.
US09/835,4611988-09-152001-04-16Antibodies having modified carbohydrate content and methods of preparation and useAbandonedUS20020028486A1 (en)

Priority Applications (1)

Application NumberPriority DateFiling DateTitle
US09/835,461US20020028486A1 (en)1988-09-152001-04-16Antibodies having modified carbohydrate content and methods of preparation and use

Applications Claiming Priority (5)

Application NumberPriority DateFiling DateTitle
US24474488A1988-09-151988-09-15
US93855792A1992-08-281992-08-28
US25152994A1994-05-311994-05-31
US08/458,666US6218149B1 (en)1988-09-151995-06-02Antibodies having modified carbohydrate content and methods of preparation and use
US09/835,461US20020028486A1 (en)1988-09-152001-04-16Antibodies having modified carbohydrate content and methods of preparation and use

Related Parent Applications (1)

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US08/458,666DivisionUS6218149B1 (en)1988-09-151995-06-02Antibodies having modified carbohydrate content and methods of preparation and use

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US20020028486A1true US20020028486A1 (en)2002-03-07

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US08/458,666Expired - LifetimeUS6218149B1 (en)1988-09-151995-06-02Antibodies having modified carbohydrate content and methods of preparation and use
US09/835,461AbandonedUS20020028486A1 (en)1988-09-152001-04-16Antibodies having modified carbohydrate content and methods of preparation and use

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US08/458,666Expired - LifetimeUS6218149B1 (en)1988-09-151995-06-02Antibodies having modified carbohydrate content and methods of preparation and use

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EP (1)EP0359096B1 (en)
JP (1)JPH02231097A (en)
AT (1)ATE159982T1 (en)
DE (1)DE68928427T2 (en)

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ATE159982T1 (en)1997-11-15
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US6218149B1 (en)2001-04-17
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DE68928427D1 (en)1997-12-11
JPH02231097A (en)1990-09-13

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