Bovine fusion antibodies

• US 9,644,021 B2
• Filed: 01/10/2014
• Issued: 05/09/2017
• Est. Priority Date: 01/11/2013
• Status: Active Grant

• Current Assignee: The Scripps Research Institute
• Original Assignee: The California Institute For Biomedical Research
• Inventors: Zhang, Yong, Schultz, Peter G., Wang, Feng
• Primary Examiner(s): Duffy, Brad
• Assistant Examiner(s): Moseley, II, Nelson B
• Application Number:
  US14/152,441
• Publication Number:
  US 20140227267A1
• Time in Patent Office: 1,215 Days
• Field of Search: None
• CPC Class Codes: C07K 14/575 Hormones derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin C07K14/665, e.g. corticotropin C07K14/695
• CPC Class Codes: C07K 14/5759 Products of obesity genes, e.g. leptin, obese (OB), tub, fat
• CPC Class Codes: C07K 14/605 Glucagons
• CPC Class Codes: C07K 14/61 Growth hormone [GH], i.e. somatotropin
• CPC Class Codes: C07K 14/635 Parathyroid hormone, i.e. parathormone; Parathyroid hormone-related peptides
• CPC Class Codes: C07K 14/64 Relaxins
• CPC Class Codes: C07K 14/811 Serine protease (E.C. 3.4.21) inhibitors
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 16/2869 against hormone receptors for antibodies against neuromediator receptors C07K16/286
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: C07K 2318/10 Immunoglobulin or domain(s) thereof as scaffolds for inserted non-Ig peptide sequences, e.g. for vaccination purposes
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: C07K 7/08 having 12 to 20 amino acids gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68

Abstract

Disclosed herein are immunoglobulin constructs comprising at least one immunoglobulin domain or fragment thereof; and a therapeutic polypeptide or derivative or variant thereof attached to or inserted into said immunoglobulin domain. Also provided are immunoglobulin constructs comprising a mammalian immunoglobulin heavy chain comprising at least a portion of a knob domain in the complementarity-determining region 3 (CDR3H) or fragment thereof; and a therapeutic polypeptide attached to or inserted into said knob domain of the CDR3H. Also provided are immunoglobulin constructs comprising a mammalian immunoglobulin heavy chain comprising at least a portion of a stalk domain in the complementarity-determining region 3 (CDR3H) or fragment thereof; and a therapeutic polypeptide attached to or inserted into said stalk domain of the CDR3H. Also described herein are methods and compositions comprising the immunoglobulin constructs described herein for treatment and prevention of a disease or condition in a subject.

Claims

1. An antibody fusion comprising:

(a) a first antibody sequence;

(b) at least a portion of an ultralong complementarity determining region 3 (CDR3), wherein the portion of the ultralong CDR3 is 15 amino acids to 61 amino acids long; and

(c) a non-antibody sequence, wherein the non-antibody sequence is selected from a polypeptide selected from a group consisting of a Mamba1, a oxyntomodulin, a 550 peptide, an Amgen 1, and a parathyroid hormone;

wherein the portion of the ultralong CDR3 is fused or inserted into a hypervariable region of the first antibody sequence, and wherein the non-antibody sequence is fused or inserted into the portion of the ultralong CDR3.

2. The antibody fusion of claim 1, wherein the first antibody sequence is selected from a bovine antibody, a human antibody, a human engineered antibody, a humanized antibody, and a chimeric antibody.

3. The antibody fusion of claim 1, wherein the ultralong CDR3 is selected from a BLV1H12 ultralong CDR3, BLV5B8 ultralong CDR3, or BF1H1 ultralong CDR3.

4. The antibody fusion of claim 1, wherein the first antibody sequence comprises a heavy chain sequence.

5. The antibody fusion of claim 1, wherein the first antibody sequence comprises an amino acid sequence that is at least 90% homologous to SEQ ID NO:

9 or is encoded by a nucleotide sequence that is at least 90% homologous to SEQ ID NO;

2.

6. The antibody fusion of claim 1, wherein the ultralong CDR3 is selected from a bovine ultralong CDR3, a camelid ultralong CDR3, and a shark ultralong CDR3.

7. The antibody fusion of claim 1, wherein the ultralong CDR3 is a bovine ultralong CDR3.

8. The antibody fusion of claim 1, wherein the antibody fusion comprises an amino acid sequence that is at least 90% homologous to SEQ ID NOs:

10-14 or is encoded by a nucleotide sequence that is at least 90% homologous to SEQ ID NOs;

3-7.

9. The antibody fusion of claim 1, further comprising a second antibody sequence.

10. The antibody fusion of claim 9, wherein the second antibody sequence comprises an amino acid sequence that is at least 90% homologous to SEQ ID NO:

8 or is encoded by a nucleotide sequence that is at least 90% homologous to SEQ ID NO;

1.

11. The antibody fusion of claim 1, further comprising a first linker sequence.

12. The antibody fusion of claim 11, further comprising a second linker sequence.

13. The antibody fusion of claim 12, wherein the first linker sequence attaches the non-antibody sequence to the first antibody sequence, the second linker sequence attaches the non-antibody sequence to the first antibody sequence, or both the first and second linker sequences attach the non-antibody sequence to the first antibody sequence.

14. The antibody fusion of claim 12, wherein the first linker sequence comprises a sequence selected from SEQ ID NOs:

15-17, 103, and 129, the second linker sequence comprises a sequence selected from SEQ ID NOs;

15-17, 103, and 129, or both the first and second linker sequences each independently comprise a sequence selected from SEQ ID NOs;

15-17, 103, and 129.

15. The antibody fusion of claim 1, further comprising one or more proteolytic cleavage sites.

16. The antibody fusion of claim 15, wherein the one or more proteolytic cleavage sites comprises SEQ ID NO:

104.

17. A vector comprising a polynucleotide comprising a sequence that is at least 90% homologous to SEQ ID NOs:

3-7 or a sequence that encodes for an amino acid that is at least 90% homologous to SEQ ID NOs;

10-14.

18. A host cell comprising a polynucleotide comprising a sequence that is at least 90% homologous to SEQ ID NOs:

3-7 or expressing an amino acid that is at least 90% homologous to SEQ ID NOs;

10-14.

19. A method of producing an antibody fusion, the method comprising culturing the host cell of claim 18 under conditions wherein the host cell expresses the polynucleotide comprising the sequence that is at least 90% homologous to SEQ ID NOs:

3-7 or the host cell produces the amino acid that is at least 90% homologous to SEQ ID NOs;

10-14.