CTLA4-Ig immunoadhesins
The present application relates to CTLA4-Ig immunoadhesins that target CD80 and CD86, and their use, particularly for therapeutic purposes.
1. A nucleic acid encoding an immunoadhesin, wherein said immunoadhesin comprises a first domain comprising a variant CTLA4 as compared to SEQ ID NO:2 and a second domain comprising an IgG Fc region, wherein said CTLA4 variant comprises an amino acid modification selected from the group consisting of T51N, A29H, M53Y, L61E, and K93Q, wherein said variant provides enhanced binding to B7-1, B7-2, or both B7-1 and B7-2, and wherein amino acid numbering is according to the EU index.
2. A nucleic acid of claim 1 , wherein said nucleic acid encodes an amino acid sequence selected from the group consisting of SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO: 37, or SEQ ID NO: 38.
3. A nucleic acid of claim 1 , wherein said nucleic acid encodes a CTLA4 variant comprising a combination of substitutions selected from the group consisting of A29H/K93Q, A29H/M53Y, A29H/T51N, T51N/K93Q, T51N/M53Y, A29H/L61E/K93Q, A29H/M53Y/K93Q, A29H/M53Y/L61E, A29H/T51N/L61E, M53Y/L61E/K93Q, T51N/L61E/K93Q, T51N/M53Y/L61E, A29H/M53Y/L61E/K93Q, A29H/T51N/L61E/K93Q, A29H/T51N/M53Y/K93Q, A29H/T51N/M53Y/L61E, T51N/M53Y/L61E/K93Q, and A29H/T51N/M53Y/L61E/K93Q.
4. A nucleic acid of claim 3 , wherein said nucleic acid encodes a CTLA4 variant comprising a combination of substitutions selected from the group consisting of T51N/L61E/K93Q, A29H/T51N/L61E/K93Q, A29H/T51N, T51N/M53Y, and T51N/M53Y/L61E.
5. A nucleic acid of claim 4 , wherein said nucleic acid encodes a CTLA4 variant comprising substitutions A29H/T51N/L61E/K93Q.
6. A nucleic acid of claim 4 , wherein said nucleic acid encodes a CTLA4 variant comprising substitutions T51N/L61E/K93Q.
7. A nucleic acid of claim 4 , wherein said nucleic acid encodes a CTLA4 variant comprising substitutions A29H/T51N.
8. A nucleic acid of claim 4 , wherein said nucleic acid encodes a CTLA4 variant comprising substitutions T51N/M53Y.
9. A nucleic acid of claim 4 , wherein said nucleic acid encodes a CTLA4 variant comprising substitutions T51N/M53Y/L61E.
10. An expression vector comprising the nucleic acid of claim 1 .
11. A cultured host cell comprising a nucleic acid encoding an immunoadhesin, wherein said immunoadhesin comprises a first domain comprising a variant CTLA4 as compared to SEQ ID NO:2 and a second domain comprising an IgG Fc region, wherein said CTLA4 variant comprises an amino acid modification selected from the group consisting of T51N, A29H, M53Y, L61E, and K93Q, wherein said variant provides enhanced binding to B7-1, B7-2, or both B7-1 and B7-2, and wherein amino acid numbering is according to the EU index.
12. The cultured host cell according to claim 11 , wherein said immunoadhesin comprises a CTLA4 variant comprising substitutions A29H/T51N/L61E/K93Q.
13. A cultured host cell comprising an expression vector comprising a nucleic acid encoding an immunoadhesin, wherein said immunoadhesin comprises a first domain comprising a variant CTLA4 as compared to SEQ ID NO:2 and a second domain comprising an IgG Fc region, wherein said CTLA4 variant comprises an amino acid modification selected from the group consisting of T51N, A29H, M53Y, L61E, and K93Q, wherein said variant provides enhanced binding to B7-1, B7-2, or both B7-1 and B7-2, and wherein amino acid numbering is according to the EU index.
14. The cultured host cell according to claim 13 , wherein said immunoadhesin comprises a CTLA4 variant comprising substitutions A29H/T51N/L61E/K93Q.
15. A method of making an immunoadhesin comprising a) culturing the cultured host cell of claim 11 under conditions wherein said immunoadhesin is produced; and b) purifying said immunoadhesin.
16. A method of making an immunoadhesin comprising a) culturing the cultured host cell of claim 13 under conditions wherein said immunoadhesin is produced; and b) purifying said immunoadhesin.