Materials and methods for performing histochemical assays for human pro-epiregulin and amphiregulin

Patent metadata

Field	Value
Publication number	US-11747339-B2
Application number	US-202016949252-A
Country	US
Kind code	B2
Filing date	Oct 21, 2020
Priority date	Jun 29, 2015
Publication date	Sep 5, 2023
Grant date	Sep 5, 2023

Abstract

The invention provides anti-human pro-epiregulin and anti-human amphiregulin antibodies and methods of using the same. Anti-EREG antibodies raised against amino acids 148-169 and 156-169 of the human EREG protein, and anti-AREG antibodies raised against amino acids 238-252 of the human AREG protein are disclosed. Methods of using these antibodies to detect EREG and AREG and kits and other products for performing such methods are also disclosed.

First claim

The invention claimed is:

1. An antibody or antibody fragment capable of specifically binding to amino acids 156-169 of SEQ ID NO: 1, wherein the antibody comprises a heavy chain comprising the following hypervariable regions (HVRs):
(a) an HVR-H1 comprising SEQ ID NO: 18;
(b) an HVR-H2 comprising SEQ ID NO: 19; and
(c) an HVR-H3 comprising SEQ ID NO: 20; and

wherein the antibody or antibody fragment further comprises a light chain comprising the following HVRs:
(h) an HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 25 and SEQ ID NO: 26;
(i) an HVR-L2 comprising SEQ ID NO: 27; and
(j) an HVR-L3 comprising SEQ ID NO: 28.

2. The antibody or antibody fragment of claim 1 , wherein the heavy chain further comprises the following variable domain framework regions (FRs):
(d) FR-H1 comprising SEQ ID NO: 21;
(e) FR-H2 comprising SEQ ID NO: 22;
(f) FR-H3 comprising SEQ ID NO: 23; and
(g) FR-H4 comprising SEQ ID NO: 24.

3. The antibody or antibody fragment of claim 1 , wherein the light chain further comprises the following variable domain framework regions (FRs):
(k) FR-L1 comprising SEQ ID NO: 29;
(l) FR-L2 comprising SEQ ID NO: 30;
(m) FR-L3 comprising SEQ ID NO: 31; and
(n) FR-L4 comprising SEQ ID NO: 32.

4. The antibody or antibody fragment of claim 1 , wherein the antibody or antibody fragment comprises
a heavy chain variable region (VH) comprising an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 33.

5. The antibody or antibody fragment of claim 4 , wherein the VH comprises SEQ ID NO: 33.

6. The antibody of claim 1 , wherein the antibody is a monoclonal antibody.

7. The antibody of claim 6 , wherein the monoclonal antibody is a rabbit monoclonal antibody.

8. The antibody of claim 6 , wherein the monoclonal antibody is an IgG antibody.

9. The antibody fragment of claim 1 , wherein the antibody fragment is selected from the group consisting of Fab, single chain variable fragment (scFv), Fv, Fab′, Fab′-SH, F(ab′)2, and diabody.

10. The antibody or antibody fragment of claim 1 , wherein the antibody or antibody fragment comprises a light chain variable region (VL) comprising an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 34 or SEQ ID NO: 35.

11. The antibody or antibody fragment of claim 4 , wherein the antibody or antibody fragment comprises a light chain variable region (VL) comprising an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 34.

12. The antibody or antibody fragment of claim 11 , wherein the VH comprises SEQ ID NO: 33 and the VL comprises SEQ ID NO: 34.

13. The antibody or antibody fragment of claim 4 , wherein the antibody or antibody fragment comprises a light chain variable region (VL) comprising an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 35.

14. The antibody or antibody fragment of claim 13 , wherein the VH comprises SEQ ID NO: 33 and the VL comprises SEQ ID NO: 35.

15. A method of detecting the presence or expression level of human pro-epiregulin in a biological sample comprising:
contacting the biological sample with the antibody or antibody fragment of claim 1 ; and
detecting the presence of the bound antibody or antibody fragment.

16. The method of claim 15 , wherein the detecting is by immunohistochemistry, immunofluorescence, or immunoblot.

17. The method of claim 15 , wherein the biological sample comprises a fixed tissue.

18. The method of claim 17 , wherein the fixed tissue is a formalin-fixed paraffin embedded (FFPE) tissue.

19. The method of claim 15 , wherein the biological sample is from a subject having or predisposed to cancer.

20. The method of claim 19 , wherein the cancer is colon cancer, breast cancer, or lung cancer.

Assignees

• Ventana Med Syst Inc

Inventors

• Couto Fernando Jose Rebelo Do
• Liao Zhiming
• Muranyi Andrea
• Shanmugam Kandavel
• Singh Shalini
• Zhu Yifei

Classifications

• G01N33/57595 Primary

  involving intracellular compounds · CPC title

• A01B39/18

  for weeding · CPC title

• G01N33/57496 Primary

  Physics · mapped topic

• C07K16/22

  against growth factors {; against growth regulators} · CPC title

• C07K16/30

  from tumour cells · CPC title