Modular assembly of antibody genes, antibodies prepared thereby and use

• US 5,698,435 A
• Filed: 06/06/1995
• Issued: 12/16/1997
• Est. Priority Date: 11/01/1985
• Status: Expired due to Term

• Current Assignee: Xoma Technology Ltd. (Xoma Corp.)
• Original Assignee: Xoma Corp.
• Inventors: Horwitz, Arnold H., Robinson, Randy R., Lei, Shau-Ping, Liu, Alvin Y., Wall, Randolph, Better, Marc, Wilcox, Gary L.
• Primary Examiner(s): Ziska, Suzanne E.
• Application Number:
  US08/467,140
• Time in Patent Office: 924 Days
• Field of Search: 435/69.1, 435/172.3, 435/69.3, 435/240.2
• US Class Current: 435/328
• 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: C07K 14/43 Sweetening agents, e.g. thaumatin, monellin
• CPC Class Codes: C07K 16/3046 Stomach, Intestines
• CPC Class Codes: C07K 16/462 Igs containing a variable region (Fv) from one specie and a constant region (Fc) from another
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/732 Antibody-dependent cellular cytotoxicity [ADCC]
• CPC Class Codes: C07K 2317/734 Complement-dependent cytotoxicity [CDC]
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/02 containing a signal sequence
• CPC Class Codes: C07K 2319/034 containing a motif for targeting to the periplasmic space of Gram negative bacteria as a soluble protein, i.e. signal sequence should be cleaved
• CPC Class Codes: C07K 2319/036 targeting to the medium outside of the cell, e.g. type III secretion
• 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: C12N 15/70 Vectors or expression systems specially adapted for E. coli
• CPC Class Codes: C12N 15/81 for yeasts
• CPC Class Codes: C12N 9/88 Lyases (4.)

Abstract

The invention relates to cDNA genetic sequences, vehicles containing same as well as hosts transformed therewith, for the production of chimeric immunoglobulin molecules, functional fragments thereof and immunoglobulin derivatives exhibiting novel functional properties comprising human constant region modules and non-human variable region modules, or for class switching antibody molecules and/or chains. The invention also relates to DNA coding for pectate lyase signal peptide has been cloned on a plasmid to create a secretion vector which is capable of producing a chosen protein which is transported across the bacterial membrane. The secretion vector has been used to secrete extracellular thaumatin and extracellular chimeric antibody fragments. The proteins produced by this vector have biological activity. The thaumatin is properly folded and the antibody fragments are capable of binding antigens on target cancer cells. The invention also relates to the secretion of chimeric antibodies and fragments thereof from yeast in functional form.

Claims

1. A polynucleotide molecule encoding a secretable immunoglobulin chain, said polynuclcotide molecule comprising DNA encoding a prokaryotic secretion signal peptide directly linked to DNA encoding a member of the group consisting of an immunoglobulin light chain, an immunoglobulin light chain fragment, and an immunoglobulin heavy chain fragment, wherein said fragment comprises the entire variable region of said immunoglobulin chain.

2. The polynucleotide molecule of claim 1, wherein said immunoglobulin chain is a light chain or light chain fragment.

3. The polynucleotide molecule of claim 1, wherein said immunoglobulin chain is a heavy chain fragment.

4. The polynucleofide molecule of claim 3, wherein said heavy chain fragment is Fd.

5. The polynucleotide molecule of claim 1, wherein said heavy chain fragment, said light chain or said light chain fragment is chimeric.

6. The polynucleofide molecule of claim 1, wherein said polynucleotide molecule is dicistronic and encodes

(a) said immunoglobulin heavy chain fragment and

(b) said immunoglobulin light chain or immunoglobulin light chain fragment.

7. A vector comprising the polynucleotide molecule of any one of claims 1-6.

8. The vector of claim 7, wherein said vector is a plasmid.

9. The vector of claim 8, wherein said vector is a bacteriophage.

10. A host cell transformed with the vector of any one of claims 7-9.

11. The host cell of claim 10, wherein said host cell is a prokaryote.

12. The host cell of claim 11, wherein said pmkaryote is E. coli.

16. The host cell of any one of claims 10-12, wherein said immunoglobulin fragment is capable of being recovered from a culture supernatant of said prokaryotic host cell.

17. The prokaryotic host cell of any one of claims 10-12, wherein said immunoglobulin fragment is capable of being recovered fnma the periplasm of said prokaryotic host cell.

13. A host cell transformed with DNA encoding a prokaryotic secretion signal peptide directly linked to DNA encoding an immunoglobulin light chain or an immunoglobulin light chain fragment, and with DNA encoding a prokaryotic secretion signal peptide directly linked to DNA encoding an immunoglobulin heavy chain fragment, wherein said fragment comprises the entire variable region of said immunoglobulin chain.

14. The host cell of claim 13, wherein said DNA is encoded on a single vector.

15. The host cell of claim 13, wherein said DNA is encoded on different vectors.

18. The host cell of any one of claims 13-15, wherein said vector is selected from the group consisting of a plasmid and a bacteriophage.

19. The host cell of claim 18, wherein said immunoglobulin fragment is capable of being recovered from a culture supernatant of said prokaryotic host cell.

20. The host cell of claim 18, wherein said immunoglobulin fragment is capable of being recovered from the periplasm of said prokaryofic host cell.