A33 antigen specific immunoglobulin products and uses thereof

• US 6,342,587 B1
• Filed: 10/22/1999
• Issued: 01/29/2002
• Est. Priority Date: 10/22/1999
• Status: Expired due to Fees

• Current Assignee: Ludwig Institute for Cancer Research, Ltd., Memorial Sloan-Kettering Cancer Center, The Scripps Research Institute
• Original Assignee: Ludwig Institute for Cancer Research, Ltd., The Scripps Research Institute
• Inventors: Ritter, Gerd, Old, Lloyd J., Barbas, Carlos F. III, Rader, Christoph, Welt, Sydney
• Primary Examiner(s): Huff, Sheela
• Assistant Examiner(s): HELMS, LARRY RONALD
• Application Number:
  US09/425,638
• Time in Patent Office: 830 Days
• Field of Search: 530/387.1, 530/387.3, 530/388.1, 530/388.8, 530/388.85, 530/389.1, 530/389.7, 424/130.1, 424/134.1, 424/135.1, 424/155.1, 424/156.1, 424/133.1
• US Class Current: 530/388.85
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: C07K 16/3046 Stomach, Intestines
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/55 Fab or Fab'
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]

Abstract

The invention is directed to novel CDRs and immunoglobulin products that bind to A33 antigens and methods for their use. The invention also involves a method for making humanized antibodies, using a rabbit as a host animal, and phage display library methodologies, and the antibodies themselves. The methodology is useful, for example, in generating humanized antibodies against molecules associated with cancer, such as A33, which is associated with colon cancer.

Claims

1. A substantially pure immunoglobulin molecule which binds specifically to A33 antigen, comprising six CDRs, the amino acid sequences of which are selected from the group consisting of:

(i) SEQ ID NO.;

68 or SEQ ID NO.;

69 (V_LCDR1);

(ii) SEQ ID NO.;

70 or SEQ ID NO.;

71 (V_LCDR2);

(iii) SEQ ID NO.;

72 or SEQ ID NO.;

73 (V_LCDR3);

(iv) SEQ ID NO.;

74 or SEQ ID NO.;

75 (V_HCDR1);

(v) SEQ ID NO.;

76, SEQ ID NO.;

77 or SEQ ID NO.;

78 (V_HCDR2); and

(vi) SEQ ID NO.;

79 or SEQ ID NO.;

80 (V_HCDR3),

wherein (i), (ii), (iii), (iv), (v), and (vi) are present in said immunoglobulin molecule.

2. The substantially pure immunoglobulin molecule of claim 1 wherein said immunoglobulin molecule is an antibody, an Fv fragment, an Fab fragment, a F(ab)₂ fragment, a single chain antibody, or a multimeric antibody.

3. The substantially pure immunoglobulin molecule of claim 1, wherein said immunoglobulin molecule is an IgM, IgD, IgG, IgA, or IgE molecule.

4. The substantially pure immunoglobulin molecule of claim 1, wherein said immunoglobulin molecule is a rabbit-human chimera.

5. The substantially pure immunoglobulin molecule of claim 1, wherein said immunoglobulin molecule has an affinity for A33 antigen greater than 500 pM.

6. The substantially pure immunoglobulin molecule of claim 1, wherein said immunoglobulin molecule has an affinity for A33 greater than 100 pM.

7. The substantially pure immunoglobulin molecule of claim 1, the V_LCDR regions of which comprise SEQ ID NO.:

68, SEQ ID NO.;

70, and SEQ ID NO.;

72.

8. The substantially pure immunoglobulin molecule of claim 1, the V_HCDR regions of which comprise SEQ ID NO.:

74, SEQ ID NO.;

76, and SEQ ID NO.;

79.

9. The substantially pure immunoglobulin molecule of claim 1, the V_LCDR regions of which comprise SEQ ID NOS.:

68, 70, and 72, and the V_HCDR regions which comprise SEQ ID NOS.;

74, 76, and 79.

10. The substantially pure immunoglobulin molecule of claim 1, further comprising V_LFR1, the amino acid sequence of which is selected from the group consisting of SEQ ID NO.:

81, 82, 83 or 84;

V_LFR2, the amino acid sequence of which is selected from the group consisting of SEQ ID NO.;

85, 86, 87, 88, 89, 90 or 91;

V_LFR3, the amino acid sequence of which is set forth at SEQ ID NO.;

92, 93, 94, 95 or 96; and

V_LFR4, the amino acid sequence of which is set forth at SEQ ID NO.;

97 or 98.

11. The substantially pure immunoglobulin molecule of claim 1, further comprising V_HFR1, the amino acid sequence of which is selected from the group consisting of SEQ ID NOS.:

99, 100, 101, 102, 103 and 104;

V_HFR2, the amino acid sequence of which is selected from the group consisting of SEQ ID NOS.;

105, 106, and 107;

V_HFR3, the amino acid sequence of which is selected from the group consisting of SEQ ID NOS.;

108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, and 124; and

V_HFR4, the amino acid sequence of which is selected from the group consisting of SEQ ID NOS.;

125 and 126.

13. The substantially pure immunoglobulin molecule of claim 1, the light chain of which is selected from the group consisting of SEQ ID NOS.:

20, 21, 22, 39, 40, 41, 42, 43, 44 and 45, and the heavy chain of which is selected from the group consisting of SEQ ID NOS.;

46, 47, 48, 49, 50, 51 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66 and 67.

14. The substantially pure immunoglobulin molecule of claim 13, the light chain of which is selected from the group consisting of SEQ ID NOS.:

39, 40, 41, 42, 43, 44 and 45, and the heavy chain of which is selected from the group consisting of SEQ ID NOS.;

49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66 and 67.

15. The substantially pure immunoglobulin molecule of claim 13 having (i) the light:

chain set forth in SEQ ID NO.;

39 and the heavy chain set forth in SEQ ID NO.;

50, (ii) the light chain set forth in SEQ ID NO.;

40 and the heavy chain set forth in SEQ ID NO.;

51, (iii) the light chain set forth in SEQ ID NO.;

41 and the heavy chain set forth in SEQ ID NO.;

52, (iv) the light chain set forth in SEQ ID NO.;

42 and the heavy chain set forth in SEQ ID NO.;

53, (v) the light chain set forth in SEQ ID NO.;

43 and the heavy chain set forth in SEQ ID NO.;

54, and (vi) the light chain set forth in SEQ ID NO.;

44 and the heavy chain set forth in SEQ ID NO.;

55.

12. A substantially pure immunoglobulin molecule which binds specifically to A33 antigen, comprising;

V_LCDR1;

SEQ ID NO.;

68

V_LCDR2;

SEQ ID NO.;

70

V_LCDR3;

SEQ ID NO.;

72

V_HCDR1;

SEQ ID NO.;

74

V_HCDR2;

SEQ ID NO.;

76

V_HCDR3;

SEQ ID NO.;

79

V_LFR1;

SEQ ID NO.;

81

V_LFR2;

SEQ ID NO.;

87

V_LFR3;

SEQ ID NO.;

92

V_LFR4;

SEQ ID NO.;

97

V_HFR1;

SEQ ID NO.;

101

V_HFR2;

SEQ ID NO.;

107

V_HFR3;

SEQ ID NO.;

110, and

V_HFR4;

SEQ ID NO.;

126.