Recombinant yeast transformant and process for preparing immunoglobulin Fc fragment employing the same

Patent metadata

Field	Value
Publication number	US-9394546-B2
Application number	US-201414763722-A
Country	US
Kind code	B2
Filing date	Feb 3, 2014
Priority date	Jan 31, 2013
Publication date	Jul 19, 2016
Grant date	Jul 19, 2016

Abstract

The present invention relates to a transformant prepared by introducing an expression vector comprising a polynucleotide encoding for a human immunoglobulin Fc fragment into Pichia sp. yeast, a method for producing an immunoglobulin Fc fragment comprising culturing the transformant, and recovering the immunoglobulin Fc fragment from the culture, and an immunoglobulin Fc fragment, prepared by the above method for use as a drug carrier. The transformant is suggested as a solution to the problems associated with the use of E. coli or animal cells as hosts for producing immunoglobulin Fc fragments useful as drug carriers, so that it can find various applications in the effective and economical production of drugs.

First claim

The invention claimed is:

1. A transformant, modified by introducing an expression vector comprising a polynucleotide encoding a human immunoglobulin Fc fragment into Pichia sp. Yeast, wherein the expression vector is pPIC9K-IgG4Fc having the cleavage map of FIG. 1 , or pPIC9K-mPSCFc having the cleavage map of FIG. 2 .

2. The transformant of claim 1 , wherein the Pichia sp. yeast is Pichia pastoris.

3. The transformant of claim 1 , wherein the immunoglobulin is IgG.

4. The transformant of claim 3 , wherein the IgG is IgG1, IgG2, IgG3 or IgG4.

5. The transformant of claim 1 , wherein the immunoglobulin Fc fragment comprises an amino acid sequence of SEQ ID NO: 10 or SEQ ID NO: 11.

6. A transformant, modified by introducing an expression vector comprising a polynucleotide encoding a human immunoglobulin Fc fragment into Pichia sp. yeast, wherein the polynucleotide comprises a nucleotide sequence of SEQ ID NO: 1 or SEQ ID NO: 2.

7. A transformant, modified by introducing an expression vector comprising a polynucleotide encoding a human immunoglobulin Fc fragment into Pichia sp. yeast, wherein the transformant is Pichia ( Komagataella ) pastoris HMC041 (Accession No. KCCM11348P) or Pichia ( Komagataella ) pastoris HMC042 (Accession No. KCCM11350P).

8. A method for producing an immunoglobulin Fc fragment, comprising:
(a) culturing the transformant of claim 1 ; and
(b) recovering the immunoglobulin Fc fragment from the culture.

9. The method of claim 8 , wherein the method is characterized by no requiring an additional protein refolding process.

10. A method for producing an immunoglobulin Fc fragment, comprising:
(a) culturing the transformant of claim 6 ; and
(b) recovering the immunoglobulin Fc fragment from the culture.

11. The method of claim 10 , wherein the method is characterized by no requiring an additional protein refolding process.

12. A method for producing an immunoglobulin Fc fragment, comprising:
(a) culturing the transformant of claim 7 ; and
(b) recovering the immunoglobulin Fc fragment from the culture.

13. The method of claim 12 , wherein the method is characterized by no requiring an additional protein refolding process.

Assignees

• Hanmi Pharm Ind Co Ltd
• Hanmi Pharm Ind Co Ltd

Inventors

• Kim Jin-Sun
• Huh Yong Ho
• Oh Euh Lim
• Kim Min Young
• Jung Sung Youb
• Kwon Se Chang

Classifications

• C07K2317/52

  Constant or Fc region; Isotype · CPC title

• C07K16/00 Primary

  Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies · CPC title

• C12N15/81 Primary

  for yeasts · CPC title

• C12N15/815 Primary

  for yeasts other than Saccharomyces · CPC title

• C07K2317/14

  Specific host cells or culture conditions, e.g. components, pH or temperature · CPC title