Human transforming growth factor

• US 4,742,003 A
• Filed: 05/14/1985
• Issued: 05/03/1988
• Est. Priority Date: 02/17/1984
• Status: Expired due to Term

• Current Assignee: Genentech, Inc. (Roche Holding AG)
• Original Assignee: Genentech, Inc. (Roche Holding AG)
• Inventors: Derynck, Rik M. A., Goeddel, David V.
• Primary Examiner(s): Wiseman, Thomas G.
• Assistant Examiner(s): Teskin, Robin Lyn
• Application Number:
  US06/695,494
• Time in Patent Office: 1,085 Days
• Field of Search: 435/68, 435/172.3, 435/317, 435/70, 435/255, 260/112 R, 260/112 B, 536/27, 935/13, 935/27, 935/48, 935/29, 935/47, 935/56, 935/63, 935/73, 935/78, 514/12, 514/2, 530/324, 530/350, 530/828
• US Class Current: 435/69.4
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61L 15/32 Proteins, polypeptides; Degradation products or derivatives thereof, e.g. albumin, collagen, fibrin, gelatin A61L15/225 takes precedence
• CPC Class Codes: C07K 14/495 Transforming growth factor [TGF]
• CPC Class Codes: C07K 16/22 against growth factors ; against growth regulators
• CPC Class Codes: G01N 2333/495 Transforming growth factor [TGF]
• CPC Class Codes: G01N 33/5748 involving oncogenic proteins
• CPC Class Codes: G01N 33/57488 involving compounds identifable in body fluids
• CPC Class Codes: G01N 33/74 involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
• CPC Class Codes: Y10S 530/828 Cancer
• CPC Class Codes: Y10S 930/12 Growth hormone, growth factor other than t-cell or b-cell growth factor, and growth hormone releasing factor; related peptides

Abstract

Methods and compositions are provided for the recombinant synthesis of the tumor growth factor-α precursor and its fragments. These are useful in therapy and diagnosis, as are antibodies raised by immunization with the tumor growth factor-α precursor and its fragment.

Claims

1. An expression vector comprising a DNA sequence encoding a human TGF-α

species selected from the group of

(1) precursor TGF-α

;

(2) a biologically active fragment of precursor TGF-α

having at least about 50 residues;

or

(3) a biologically active mutant of precursor TGF-α

or a fragment of precursor TGF-α

having at least about 50 residues, wherein an amino acid residue has been inserted, substituted or deleted in or from the amino acid sequence of the precursor TGF-α

sequence or its fragment.

2. The vector of claim 1 wherein the biologically active mutant binds to a cell surface receptor for TGF-α

and is an insertional or substitutional mutant of mature TGF-α

.

3. The vector of claim 2 wherein the fragment is mature TGF-α

.

4. The vector of claim 1 wherein the mutant comprises an insertion of a heterologous polypeptide at carboxyl-terminus and/or amino terminus of mature TGF-α

.

5. The vector of claim 3 wherein a methionyl residue is added to the amino terminus of the mature TGF-α

.

6. The vector of claim 1 which is a plasmid or virus.

7. A method for producing a TGF-α

species, comprising

(a) preparing an expression vector comprising a DNA sequence encoding a human precursor TGF-α

species selected from the group of

(1) precursor TGF-α

;

(2) a biologically active fragment of precursor TGF-α

having at least about 50 residues;

or

(3) a biologically active mutant of precursor TGF-α

, or a fragment thereof having at least about 50 residues, wherein an amino acid residue has been inserted, substituted or deleted in or from the amino acid sequence of the precursor TGF-α

sequence or its fragment;

(b) transforming said host cell with the expression vector to obtain a recombinant host cell;

(c) culturing the recombinant host cell under conditions for expression of the TGF-α

species; and

(d) recovering the TGF-α

species from the host cell culture.

8. The method of claim 7 wherein the host cell is yeast.

9. The method of claim 7 wherein the TGF-α

species is mature TGF-α

.

10. The method of claim 7 wherein the TGF-α

species is recovered as insoluble aggregates and thereafter is resolubilized.

11. The method of claim 7 wherein the TGF-α

species is secreted from the host cells and is recovered from the host cell culture medium.