Anti-ganglioside antibodies and compositions

• US 8,257,703 B2
• Filed: 03/18/2010
• Issued: 09/04/2012
• Est. Priority Date: 07/15/2003
• Status: Active Grant

• Current Assignee: Chugai Seiyaku Kabushiki Kaisha (Roche Holding AG)
• Original Assignee: Chugai Seiyaku Kabushiki Kaisha (Roche Holding AG)
• Inventors: Irie, Reiko, Tsunoda, Hiroyuki, Igawa, Tomoyuki, Sekimori, Yasuo, Tsuchiya, Masayuki
• Primary Examiner(s): OUSPENSKI, ILIA I
• Application Number:
  US12/727,162
• Publication Number:
  US 20100172899A1
• Time in Patent Office: 901 Days
• Field of Search: None
• US Class Current: 424/130.1
• 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: C07K 16/3084 against tumour-associated gangliosides
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/734 Complement-dependent cytotoxicity [CDC]
• CPC Class Codes: G01N 33/6854 Immunoglobulins
• CPC Class Codes: G01N 33/686 Anti-idiotype

Abstract

IgM can be obtained in the form of a pentamer by placing the genes encoding the H, L, and J chains on the same vector to transform appropriate host cells. The gene encoding the J chain may be introduced by co-transfection. When no J chain is expressed, the IgM is produced as a hexamer. The transformants obtained according to the present invention achieve a high yield of IgM. The present invention also provides methods which enable separation and quantification of polymeric IgM.

Claims

1. A pharmaceutical composition comprising a substantially purified anti-ganglioside GM3 IgM antibody, wherein said IgM antibody

(a) has a heavy (H) chain comprising SEQ ID NO:

2 and a light (L) chain comprising SEO ID NO;

4, and

(b) is recombinantly produced in a non-lymphoid cell.

2. The pharmaceutical composition of claim 1 comprising 80% or more pentamer IgM.

3. The pharmaceutical composition of claim 1 comprising 50% or more hexamer IgM.

4. The pharmaceutical composition of claim 3, comprising 80% or more hexamer IgM.

5. The pharmaceutical composition of claim 1 comprising IgM having a hexamer/pentamer ratio of at least 1.5.

6. The pharmaceutical composition of claim 1, wherein the non-lymphoid cell is a CHO cell.

7. The pharmaceutical composition of claim 1, wherein the IgM is produced by a method comprising

(a) culturing transformed non-lymphoid cells so that they produce the IgM at a level of at least 100 mg/L or at a level of at least 35 pg/cell/day, and

(b) collecting the IgM from the culture or a supernatant of the culture.

8. The pharmaceutical composition of claim 1, wherein the substantially pure IgM is obtained by purifying the IgM from a culture supernatant obtained from culturing transformed non-lymphoid cells producing the IgM at a level of at least 100 mg/L or at a level of at least 35 pg/cell/day.

9. The pharmaceutical composition of claim 1, wherein the IgM antibody is produced in the absence of IgM J chain.

10. The pharmaceutical composition of claim 1, wherein the IgM antibody is produced in the presence of IgM J chain.

11. A pharmaceutical composition comprising a substantially purified anti-ganglioside GM2 IgM antibody, wherein said IgM antibody

(a) has a heavy (H) chain comprising SEQ ID NO:

20 and a light (L) chain comprising SEQ ID NO;

22, and

(b) is recombinantly produced in a non-lymphoid cell.

12. The pharmaceutical composition of claim 11 comprising 80% or more pentamer IgM.

13. The pharmaceutical composition of claim 11 comprising 50% or more hexamer IgM.

14. The pharmaceutical composition of claim 13, comprising 80% or more hexamer IgM.

15. The pharmaceutical composition of claim 11 comprising IgM having a hexamer/pentamer ratio of at least 1.5.

16. The pharmaceutical composition of claim 11, wherein the non-lymphoid cell is a CHO cell.

17. The pharmaceutical composition of claim 11, wherein the IgM is produced by a method comprising

(a) culturing transformed non-lymphoid cells so that they produce the IgM at a level of at least 100 mg/L or at a level of at least 35 pg/cell/day, and

(b) collecting the IgM from the culture or a supernatant of the culture.

18. The pharmaceutical composition of claim 11, wherein the substantially pure IgM is obtained by purifying the IgM from a culture supernatant obtained from culturing transformed non-lymphoid cells producing the IgM at a level of at least 100 mg/L or at a level of at least 35 pg/cell/day.

19. The pharmaceutical composition of claim 11, wherein the IgM antibody is produced in the absence of IgM J chain.

20. The pharmaceutical composition of claim 11, wherein the IgM antibody is produced in the presence of IgM J chain.