Methods of Affinity Maturing Antibodies

• US 20110092372A1
• Filed: 10/14/2010
• Published: 04/21/2011
• Est. Priority Date: 10/14/2009
• Status: Active Grant

• Current Assignee: Janssen Biotech, Inc. (Johnson & Johnson)
• Original Assignee: Janssen Biotech, Inc. (Johnson & Johnson)
• Inventors: ALMAGRO, JUAN CARLOS, FRANSSON, JOHAN, RAGHUNATHAN, GOPALAN
• Granted Patent: US 8,580,714 B2
• Time in Patent Office: Days
• US Class Current: 506/1
• 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 16/248 IL-6
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Abstract

The invention relates to methods of affinity maturing antibodies.

Claims

1. A method of affinity maturing an antibody, comprising the steps of:

a. obtaining an amino acid sequence of the antibody light chain variable region (VL) or the antibody heavy chain variable region (VH);

b. identifying rSDRU residues in the amino acid sequence of the antibody VL or VH;

c. selecting a subset of the rSDRU residues to be diversified;

d. selecting a set of amino acids or a universal set of amino acids used for diversifying the subset of the rSDRU residues;

e. preparing a library of antibody VL or VH variants by diversifying the subset of the rSDRU residues selected in step c. with the set of amino acids selected in step d.;

f. expressing the library of antibody VH or VL variants in a host or translating the library of antibody VH or VL variants in vitro; and

g. selecting from the library of antibody VH or VL variants one or more affinity matured antibody having an improved affinity to an antigen.

2. The method of claim 1, wherein the subset of the rSDRU residues comprises residues having rSDRU>

5, rSDRU>

15, rSDRU>

30, rSDRU>

40, or rSDRU>

60.

3. The method of claim 1, wherein the subset of the rSDRU residues comprises residues having rSDRU>

40.

4. The method of claim 1, wherein the subset of the rSDRU residues reside in the antibody VH.

5. The method of claim 1, wherein the subset of the rSDRU residues reside in the antibody VL.

6. The method of claim 1, wherein the rSDRU residues comprise the antibody VL Chothia residues 32, 50, 91, 92, 93, 94, and 96.

7. The method of claim 1, wherein the rSDRU residues comprise the antibody VL Chothia residues 30a, 30c, 30d, 30f, 32, 50, 91, 92, 93, 94, and 96.

8. The method of claim 1, wherein Trp, Asn or Ser is removed from the set of amino acids or from the universal set of amino acids.

9. The method of claim 1, wherein the universal set of amino acids is RNDGHSWY, RNDGHSY, RNDGHWY, or RDGHSWY.

10. The method of claim 1, wherein the set of amino acids comprise

a. RNDGHSY at VL Chothia residue 30;

b. RNDGHWY at VL Chothia residue 30a;

c. RNDGHSY at VL Chothia residue 30c;

d. RNDGHWY at VL Chothia residue 30f;

e. RNDY at VL Chothia residue 32;

f. YWNK at VL Chothia residue 50;

g. SYGH at VL Chothia residue 91;

h. SYGN at VL Chothia residue 92;

i. STER at VL Chothia residue 93;

j. YSHT at VL Chothia residue 94; and

k. YRWL at VL Chothia residue 96.