Method for generating antibodies

• US 20050043517A1
• Filed: 08/20/2003
• Published: 02/24/2005
• Est. Priority Date: 08/20/2003
• Status: Abandoned Application

• Current Assignee: Centocor Incorporated (Johnson & Johnson)
• Original Assignee: Centocor Incorporated (Johnson & Johnson)
• Inventors: Giles-Komar, Jill, Mbow, M. Lamine, Lamb, Roberta A.
• Application Number:
  US10/644,308
• Publication Number:
  US 20050043517A1
• Time in Patent Office: Days
• US Class Current: 530/388.100
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 16/44 against material not provided for elsewhere , e.g. haptens, metals, DNA, RNA, amino acids

Abstract

Methods for generating antibodies in rodents are disclosed. The antibodies are useful as therapeutic agents, diagnostic agents or research reagents.

Claims

1. A method for generating monoclonal antibodies in a rodent comprising the steps of:

a) administering a dendritic cell expansion agent to the rodent;

b) administering a dendritic cell maturation agent to the rodent;

c) immunizing the rodent with an antigen; and

d) isolating antigen-specific antibodies.

2. The method of claim 1 wherein the dendritic cell expansion agent is Flt3 ligand (Flt3L).

3. The method of claim 2 wherein Flt3L is administered in combination with another dendritic cell expansion agent.

5. The method of claim 1 or 4 further comprising the step of administering a CD40 agonist post-immunization.

6. The method of claim 1 or 4 wherein the dendritic cell maturation agent is a type I interferon, tissue necrosis factor-α

, interleukin-6, prostaglandin-E2, interleukin-1α

, interleukin-1β

, interleukin-18, interleukin-12, interleukin-4, interleukin-23, interferon-γ

, granulocyte-macrophage colony-stimulating factor or dendritic cell associated maturation factor agonist monoclonal antibody.

7. The method of claim 6 wherein the dendritic cell maturation agent is adminstered singly or in combination with another dendritic cell maturation agent.

8. The method of claim 6 wherein the dendritic cell associated maturation factor agonist monoclonal antibody is anti-CD40.

9. The method of claim 6 wherein the type I interferon is interferon-α

(IFN-α

), interferon-β

(IFN-β

), IFN-δ

, IFN-α

1, IFN-α

2, IFN-α

2a, IFN-α

2b, IFN-α

4, IFN-α

II1, IFN-α

Con1, IFN-α

LE, IFN-α

Ly or IFN-β

2.

10. The method of claim 9 wherein the type I interferon is a combination of IFN-α

and IFN-α

.

11. The method of claim 1 or 4 wherein the rodent is a mouse.

12. The method of claim 1 wherein the mouse is a C57BL/6 mouse.

14. The method of claim 12 wherein the mouse is a transgenic mouse.

15. The method of claim 12 wherein the mouse is a knockout mouse.

16. The method of claim 12 wherein the mouse is a severe combined imumunodeficient mouse.

17. The method of claim 12 wherein the mouse is a recombination activation gene deficient mouse.

18. The method of claim 1 or 4 wherein the rodent is a rat.

4. A method for generating monoclonal antibodies in a rodent comprising the steps of:

a) administering a dendritic cell maturation agent to the rodent;

b) immunizing the rodent with an antigen; and

c) isolating antigen-specific antibodies.

13. The method of claim 4 wherein the mouse is a C57BL/6 mouse or a BALB/c mouse.

19. A method for generating antibodies in a C57BL/6 mouse comprising the steps of sequentially:

a) administering Flt3L to the mouse;

b) administering a combination of IFN-α and

IFN-β

to the mouse;

c) immunizing the mouse with an antigen; and

d) isolating antigen-specific antibodies.

22. The method of claim 19 or 20 wherein Flt3L is administered in an amount of about 8.8 μ

g to about 10 μ

g per day over a period of about 10 days to about 14 days.

23. The method of claim 19, 20 or 21 wherein the IFN-α

/β

combination is administered in an amount of about 10⁵ U to about 2×

10⁵ U each of IFN-α and

IFN-β

daily for about 3 days to about 5 days.

20. A method for generating antibodies in a C57BL/6 mouse comprising the steps of sequentially:

a) administering Flt3L to the mouse;

b) administering a combination of IFN-α and

IFN-β

to the mouse;

c) immunizing the mouse with an antigen;

d) administering a CD40 agonist; and

e) isolating antigen-specific antibodies.

24. The method of claim 20 or 21 wherein the CD40 agonist is an anti-CD40 antibody.

25. The method of claim 24 wherein the anti-CD40 antibody is administered in an amount of about 50 μ

g to about 100 μ

g per dose.

21. A method for generating antibodies in a BALB/c mouse comprising the steps of sequentially:

a) administering a combination of IFN-α and

IFN-β

to the mouse;

b) immunizing the mouse with an antigen;

c) administering a CD40 agonist; and

d) isolating antigen-specific antibodies.