Fibronectin type III domain based scaffold compositions, methods and uses

• US 10,040,842 B2
• Filed: 10/23/2015
• Issued: 08/07/2018
• Est. Priority Date: 10/31/2008
• Status: Active Grant

• Current Assignee: Janssen Biotech, Inc. (Johnson & Johnson)
• Original Assignee: Janssen Biotech, Inc. (Johnson & Johnson)
• Inventors: Jacobs, Steven, O'Neil, Karyn
• Primary Examiner(s): BOESEN, CHRISTIAN C
• Application Number:
  US14/921,589
• Publication Number:
  US 20160039909A1
• Time in Patent Office: 1,019 Days
• Field of Search: None
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 13/00 Drugs for disorders of the urinary system
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 33/00 Antiparasitic agents
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 39/02 Antidotes
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C07K 14/78 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
• CPC Class Codes: C40B 40/08 Libraries containing RNA or DNA which encodes proteins, e.g. gene libraries
• CPC Class Codes: C40B 40/10 Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: C40B 50/06 : Biochemical methods, e.g. using enzymes or whole viable microorganisms

Abstract

A protein scaffold based on a consensus sequence of fibronectin type III (FN3) proteins, such as the tenth FN3 repeat from human fibronectin (human Tenascin), including isolated nucleic acids that encode a protein scaffold, vectors, host cells, and methods of making and using thereof have applications in diagnostic and/or therapeutic compositions, methods and devices. In particular, protein scaffold molecules binding to IgG have been identified as useful for diagnostic and/or therapeutic applications.

Claims

1. An isolated nucleic acid molecule comprising a nucleotide sequence encoding a protein scaffold comprising the amino acid sequence of SEQ ID NO:

16.

2. An isolated nucleic acid vector comprising the isolated nucleic acid molecule of claim 1.

3. A prokaryotic or eukaryotic host cell comprising the isolated nucleic acid molecule of claim 1.

4. The host cell of claim 3, wherein said host cell is at least one selected from E. coli BL21Star(DE3), other E. coli cell, yeast, COS-1, COS-7, HEK293, BHK21, CHO, BSC-1, Hep G2, 653, SP2/0, 293, HeLa, myeloma, or lymphoma cells, or any derivative, immortalized or transformed cell thereof.

5. The isolated nucleic acid molecule of claim 1, comprising the nucleotide sequence set forth in SEQ ID NO:

17.

6. An isolated nucleic acid vector comprising the isolated nucleic acid molecule of claim 5.

7. A prokaryotic or eukaryotic host cell comprising the isolated nucleic acid molecule of claim 5.

8. The host cell of claim 7, wherein said host cell is at least one selected from E. coli BL21Star(DE3), other E. coli cell, yeast, COS-1, COS-7, HEK293, BHK21, CHO, BSC-1, Hep G2, 653, SP2/0, 293, HeLa, myeloma, or lymphoma cells, or any derivative, immortalized or transformed cell thereof.

9. An isolated nucleic acid molecule comprising a nucleotide sequence encoding a protein scaffold based on a fibronectin type III (FN3) domain comprising:

a backbone portion having an amino acid sequence identical to SEQ ID NO;

16 at residues 1-21, 29-74, and 82-89;

a B;

C loop portion between residues 21 and 29 of SEQ ID NO;

16 having at least 75% identity to residues 22-28 of the amino acid sequence of SEQ ID NO;

16; and

an F;

G loop portion between residues 74 and 82 of SEQ ID NO;

16 having at least 75% identity to residues 75-81 of the amino acid sequence of SEQ ID NO;

16.

10. An isolated nucleic acid vector comprising the isolated nucleic acid molecule of claim 9.

11. A prokaryotic or eukaryotic host cell comprising the isolated nucleic acid molecule of claim 9.

12. The host cell of claim 11, wherein said host cell is at least one selected from E. coli BL21Star(DE3), other E. coli cell, yeast, COS-1, COS-7, HEK293, BHK21, CHO, BSC-1, Hep G2, 653, SP2/0, 293, HeLa, myeloma, or lymphoma cells, or any derivative, immortalized or transformed cell thereof.