Anti-GP73 monoclonal antibodies and methods of obtaining the same

• US 9,469,686 B2
• Filed: 03/15/2013
• Issued: 10/18/2016
• Est. Priority Date: 03/15/2013
• Status: Active Grant

• Current Assignee: Abbott Laboratories
• Original Assignee: Abbott Laboratories
• Inventors: Strobel, Carolyn J., Zhao, Cheng, Tu, Bailin, Ziemann, Robert N., Tieman, Bryan C., Hemken, Philip M., Ramsay, Carol S., Hawksworth, David J., Birkenmeyer, Larry G., Brophy, Susan E., Dowell, Barry L., Muerhoff, Anthony S.
• Primary Examiner(s): PAK, MICHAEL D
• Application Number:
  US13/836,229
• Publication Number:
  US 20140271651A1
• Time in Patent Office: 1,313 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/33 Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
• CPC Class Codes: C07K 2317/34 Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: G01N 2333/47 Assays involving proteins of known structure or function as defined in the subgroups
• CPC Class Codes: G01N 2800/085 Liver diseases, e.g. portal hypertension, fibrosis, cirrhosis, bilirubin
• CPC Class Codes: G01N 2800/50 Determining the risk of developing a disease
• CPC Class Codes: G01N 33/57438 of liver, pancreas or kidney
• CPC Class Codes: G01N 33/6893 related to diseases not provided for elsewhere

Abstract

Disclosed herein are antibodies and methods of using said antibodies to detect Golgi protein 73 (GP73) and fucosylated GP73 in a sample.

Claims

1. An isolated antibody which binds to Golgi Protein 73 (“

GP73”

), wherein the antibody comprises a variable heavy domain chain comprising a CDR1 comprising the amino acid sequence of SEQ ID NO;

42, a CDR2 comprising the amino acid sequence of SEQ ID NO;

43, and a CDR comprising the amino acid sequence of SEQ ID NO;

44, and a variable light domain chain comprising a CDR1 comprising the amino acid sequence of SEQ ID NO;

46, a CDR2 comprising the amino acid sequence of SEQ ID NO;

47, and a CDR3 comprising the amino acid sequence of SEQ ID NO;

48.

2. The isolated antibody of claim 1, wherein the antibody comprises:

i) a variable heavy region comprising a sequence of SEQ ID NO;

41;

ii) a variable light region comprising a sequence of SEQ ID NO;

45;

iii) a variable heavy domain comprising the amino acid sequence of SEQ ID NO;

41 and a variable light domain region comprising the amino acid sequence of SEQ ID NO;

45.

3. The isolated antibody of claim 1, wherein the binding of the antibody to GP73 is:

i) insensitive to the presence or absence of a fucose sugar moiety on GP73.

4. The isolated antibody of claim 1, wherein said antibody has:

i) an equilibrium dissociation constant (KD) of between about 4.0×

10<sup>−

9</sup> to about 1.8×

10<sup>−

12</sup>;

ii) a dissociation rate (koff) of between about 1.5×

10<sup>−

3</sup> to about 8.0×

10<sup>−

6</sup>;

iii) an association rate (kon) of between about 1.0×

10⁵ to about 4.1×

10⁶;

or

iv) an association rate (kon) of between about 1.0×

10⁵ to about 4.1×

10₆.

5. A pharmaceutical composition comprising the antibody of claim 1.

6. A kit comprising the antibody of claim 1.

7. A kit comprising an isolated nucleic acid encoding the antibody of claim 1.

9. A kit for assaying a test sample for fucosylated GP73, which kit comprises at least one capture binding protein, wherein the capture binding protein binds to GP73 or a fragment of GP73, at least one detection binding protein, wherein the detection binding protein binds to GP73 that is not bound by the capture binding protein, and instructions for assaying the test sample for GP73, wherein the at least one capture binding protein comprises an antibody whose binding to GP73 is insensitive to the presence or absence of a fucose sugar moiety on GP73 and the at least one detection binding protein comprises an antibody whose binding to GP73 is sensitive to fucosylation of GP73, and wherein the at least one capture binding protein comprises the antibody of claim 1.

10. A kit for assaying a test sample for fucosylated GP73, which kit comprises at least one capture binding protein, wherein the capture binding protein binds to GP73 or a fragment of GP73, at least one detection binding protein, wherein the detection binding protein binds to GP73 that is not bound by the capture binding protein, and instructions for assaying the test sample for GP73, wherein the at least one capture binding protein comprises an antibody whose binding to GP73 is sensitive to fucosylation of GP73 and the at least one detection binding protein comprises an antibody whose binding to GP73 is insensitive to fucosylation of GP73, and wherein the at least one detection binding protein comprises the antibody of claim 1.

11. The kit of claim 7, further comprising a reference standard indicating a GP73 concentration in a control or calibrator.

8. A kit for assaying a test sample for GP73, which kit comprises at least one capture antibody, wherein the capture antibody binds to an epitope on GP73 or a fragment of GP73, at least one detection antibody, wherein the detection antibody binds to an epitope on GP73 that is not bound by the capture antibody, and instructions for assaying the test sample for GP73, wherein the at least one capture antibody comprises:

(a) a variable heavy domain comprising the amino acid sequence os SEQ ID NO;

41 and a variable light domain comprising the amino acid sequence of SEC ID NO;

45 ;

or

(b) a variable heavy chain comprising a CDR1 comprising the amino acid sequence of SEQ ID NO;

42, a CDR2 comprising the amino acid sequence of SEQ ID NO;

43, and a CDR3 comprising the amino acid sequence of SEQ ID NO;

44 and a variable light chain comprising a CDR1 comprising the amino acid sequence of SEQ ID NO;

46, a CDR2 comprising the amino acid sequence of SEQ ID NO;

47, and a CDR3 comprising the amino acid sequence of SEQ ID NO;

48;

wherein the at least one detection antibody comprises;

(i) variable heavy domain comprising the amino acid sequence of SEQ ID NO;

25;

(ii) a variable light domain comprising the amino acid sequence of SEQ ID NO;

29;

(iii) a variable heavy domain comprising the amino acid sequence of SEQ ID NO;

25 and a variable light domain comprising the amino acid sequence of SEQ ID NO;

29;

(iv) a variable heavy chain comprising a CDR1 comprising the amino acid sequence of SEQ ID NO;

26, a CDR2 comprising the amino acid sequence of SEQ ID NO;

27, and a CDR3 comprising the amino acid sequence of SEQ ID NO;

28;

(v) a variable light chain comprising a CDR1 comprising the amino acid sequence of SEQ ID NO;

30, a CDR2 comprising the amino acid sequence of SEQ ID NO;

31, and a CDR3 comprising the amino acid sequence of SEQ ID NO;

32;

or (vi) a variable heavy chain comprising a CDR1 comprising the amino acid sequence of SEQ ID NO;

26, a CDR2 comprising the amino acid sequence of SEQ ID NO;

27, and a CDR3 comprising the amino acid sequence of SEQ ID NO;

28 and a variable light chain comprising a CDR1 comprising the amino acid sequence of SEQ ID NO;

30, a CDR2 comprising the amino acid sequence of SEQ ID NO;

31, and a CDR3 comprising the amino acid sequence of SEQ ID NO;

32,

wherein the at least one detection antibody is optionally detectably labeled.