Overexpression of the Chaperone BIP in a Heterokaryon

• US 20100062491A1
• Filed: 01/03/2008
• Published: 03/11/2010
• Est. Priority Date: 01/05/2007
• Status: Abandoned Application

• Current Assignee: Novozymes A/S (Novozymes)
• Original Assignee: Novozymes A/S (Novozymes)
• Inventors: Lehmbeck, Jan
• Application Number:
  US12/519,480
• Publication Number:
  US 20100062491A1
• Time in Patent Office: Days
• US Class Current: 435/71.100
• 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: C12N 15/80 for fungi
• CPC Class Codes: C12P 21/02 having a known sequence of two or more amino acids, e.g. glutathione

Abstract

The present invention relates to a method for increasing the production yield of a secreted antibody or antibody fragment in a filamentous fungal host cell, comprising: recombinant expression of the antibody or antibody fragment and over-expressing a BiP chaperone protein.

Claims

1. A method for increasing the production yield of a secreted antibody or antibody fragment in a filamentous fungal host cell, comprising:

recombinant expression of the antibody or antibody fragment and over-expression of a BiP chaperone protein.

2. The method according to claim 1, wherein the BiP chaperone is an Aspergillus BiP protein.

3. The method according to claim 2, wherein the BiP chaperone protein is BiPA from Aspergillus oryzae.

4. The method according to claim 1, wherein the BiP protein expression is controlled by a promoter selected from the group consisting of A. oryzae TAKA amylase, NA2, NA2-tpi, glaA, tpi, gpd, tef1, and pgkA promoters.

5. The method according to claim 1, wherein the antibody or antibody fragment is selected from the group consisting of IgG1, IgG2, IgG3, IgG4, IgM, IgA, IgD, IgE, F(ab′

)₂, and Fab.

6. The method according to claim 1, wherein the antibody is an IgG antibody.

7. The method according to claim 1, wherein the filamentous fungal host cell has a reduced or no expression of one or more proteases selected from the group consisting of PepC, KexB, Alp, NpI.

8. The method according to claim 1, wherein the filamentous fungal host cell is a heterokaryon.

9. The method according to claim 1, wherein the filamentous fungal cell is selected from the group consisting of Acremonium, Aspergillus, Fusarium, Humicola, Mucor, Myceliophthora, Neurospora, Penicillium, Thielavia, Tolypocladium, or Trichoderma.

10. The method according to claim 9, wherein the Aspergillus cell is selected from the group consisting of Aspergillus awamori, Aspergillus foetidus, Aspergillus japonicus, Aspergillus nidulans, Aspergillus niger or Aspergillus oryzae.

11. The method according to claim 10, wherein the Aspergillus cell is A. oryzae.

12. The method according to claim 10, wherein the Aspergillus cell is A. niger.