High potency recombinant antibodies and method for producing them
High potency antibodies, including immunologically active fragments thereof, having high kinetic association rate constants and optional high affinities are disclosed, along with methods for producing such antibodies. The high potency antibodies disclosed herein are of either the neutralizing or non-neutralizing type and have specificity for antigens displayed by microorganisms, especially viruses, as well as antigenic sites present on cancer cells and on various types of toxins, and the products of toxins. Processes for producing high potency neutralizing antibodies and increasing the potency of already existing neutralizing antibodies are also described. Methods of using said antibodies in the prevention and/or treatment of diseases, especially diseases induced or caused by viruses, are disclosed.
1. A method for preventing a disease caused by respiratory syncytial virus (RSV) in a patient at risk of such disease, comprising administering to the patient an antibody that specifically binds to the F protein of a RSV, wherein the antibody:
(a) has an association rate constant (k on ) of at least 2.5×10 5 M −1 s −1 as measured by surface plasmon resonance; and
(b) comprises the following complementarity determining regions (CDRs) with one or more amino acid changes at the underlined and bolded positions in one or more of the CDRs:
heavy chain variable region
(VH) CDR1
T S GMSVG;
(SEQ ID NO: 8)
VH CDR2
DIWWD D K KD YNPSLK S ;
(SEQ ID NO: 9)
VH CDR3
S MI T N W YFDV;
(SEQ ID NO: 10)
light chain variable region
(VL) CDR1
SA SS S VGYMH;
(SEQ ID NO: 5)
VL CDR2
DT SKLA S;
(SEQ ID NO: 6)
and
VL CDR3
FQGS G YP F T;
(SEQ ID NO: 7)
and wherein at least one of said one or more amino acid changes is in VH CDR2.
2. A method for treating a disease caused by RSV in a patient afflicted with such disease, comprising administering to the patient an antibody that specifically binds to the F protein of a RSV, wherein the antibody:
(a) has a k on of at least 2.5×10 5 M −1 s −1 as measured by surface plasmon resonance; and
(b) comprises the following CDRs with one or more amino acid changes at the underlined and bolded positions in one or more of the CDRs:
VH CDR1
T S GMSVG;
(SEQ ID NO: 8)
VH CDR2
DIWWD D K KD YNPSLK S ;
(SEQ ID NO: 9)
VH CDR3
S MI T N W YFDV;
(SEQ ID NO: 10)
VL CDR1
SA SS S VGYMH;
(SEQ ID NO: 5)
VL CDR2
DT SKLA S;
(SEQ ID NO: 6)
and
VL CDR3
FQGS G YP F T;
(SEQ ID NO: 7)
and wherein at least one of said one or more amino acid changes is in VH CDR2.
3. The method of claim 1 , wherein the antibody binds to the same epitope of the RSV F antigen as an existing antibody comprising a VH having the amino acid sequence of SEQ ID NO:4 ( FIG. 2B ) and a VL having the amino acid sequence of SEQ ID NO:3 ( FIG. 2A ).
4. The method of claim 2 , wherein the antibody binds to the same epitope of the RSV F antigen as an existing antibody comprising a VH having the amino acid sequence of SEQ ID NO:4 ( FIG. 2B ) and a VL having the amino acid sequence of SEQ ID NO:3 ( FIG. 2A ).
5. The method of claim 1 , wherein the antibody has an EC 50 of less than 6.0 nM as measured in a microneutralization assay.
6. The method of claim 2 , wherein the antibody has an EC 50 of less than 6.0 nM as measured in a microneutralization assay.
7. The method of claim 1 , wherein the antibody has a k on of at least 5×10 5 M −1 s −1 as measured by surface plasmon resonance.
8. The method of claim 2 , wherein the antibody has a k on of at least 5×10 5 M −1 s −1 as measured by surface plasmon resonance.
9. The method of claim 1 , wherein the antibody has a k on of at least 7.5×10 5 M −1 s −1 as measured by surface plasmon resonance.
10. The antibody of claim 2 , wherein the antibody has a k on of at least 7.5×10 5 M −1 s −1 as measured by surface plasmon resonance.
11. The method of claim 1 , wherein the antibody has a k on of between 2.5×10 5 M −1 s −1 and 7.5×10 5 M −1 s −1 as measured by surface plasmon resonance.
12. The method of claim 2 , wherein the antibody has a k on of between 2.5×10 5 M −1 s −1 and 7.5×10 5 M −1 s −1 as measured by surface plasmon resonance.
13. The method of claim 1 , wherein the antibody has a k on of between 3.7×10 5 M −1 s −1 and 7.9×10 5 M −1 s −1 as measured by surface plasmon resonance.
14. The method of claim 2 , wherein the antibody has a k on of between 3.7×10 5 M −1 s −1 and 7.9×10 5 M −1 s −1 as measured by surface plasmon resonance.
15. The method of claim 5 , wherein the antibody has an EC 50 of less than 3.0 nM.
16. The method of claim 6 , wherein the antibody has an EC 50 of less than 3.0 nM.
17. The method of claim 5 , wherein the antibody has an EC 50 of less than 1.0 nM.
18. The method of claim 6 , wherein the antibody has an EC 50 of less than 1.0 nM.
19. The method of claim 1 , wherein the antibody is a monoclonal antibody.
20. The method of claim 2 , wherein the antibody is a monoclonal antibody.
21. The method of claim 1 , wherein the antibody is a Fab or F(ab′) 2 fragment.
22. The method of claim 2 , wherein the antibody is a Fab or F(ab′) 2 fragment.
23. The method of claim 1 , wherein the antibody is a whole tetrameric antibody molecule.
24. The method of claim 2 , wherein the antibody is a whole tetrameric antibody molecule.
25. The method of claim 1 , wherein the antibody is a humanized antibody.
26. The method of claim 2 , wherein the antibody is a humanized antibody.
27. The method of claim 1 , wherein the antibody blocks RSV-induced fusion.
28. The method of claim 2 , wherein the antibody blocks RSV-induced fusion.
29. The method of claim 2 , wherein the antibody blocks RSV-induced fusion after viral attachment to cells.
30. The method of claim 2 , wherein the antibody blocks RSV-induced fusion after viral attachment to cells.