Purification process for removal of tyrosine sulfation antibody variants; purified compositions
AssigneeMerck Sharp & Dohme Corp.
InventorsJia Zhao, Sandra Rios, Svetlana Dukleska Schussler
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Abstract
The present invention relates to purified antibody and antigen-binding fragment compositions that lack sulfated tyrosine on one or more tyrosine residues in the immunoglobulin chains. Purification methods for removing sulfated tyrosine variants from antibody and antigen-binding fragment compositions are also provided.
Claims24
A purified composition comprising non-CDR-L1 tyrosine sulfated antibodies that specifically bind to human lymphocyte activation gene 3 (anti-hLAG3) comprising:
- a heavy chain amino acid sequence of SEQ ID NO:
- 52 and a light chain amino acid sequence of SEQ ID NO:
- 51, and less than about 0.5% CDR-L1 tyrosine sulfated anti-hLAG3antibodies as compared to non-CDR-L1 tyrosine sulfated anti-hLAG3 antibodies as determined by mass spectrometry, wherein the CDR-L1 tyrosine sulfated anti-hLAG3 antibodies comprise a heavy chain amino acid sequence of SEQ ID NO:
- 52 and a light chain amino acid sequence of SEQ ID NO:
- 51 with the tyrosine at CDR-L1 sulfated,
- wherein the non-CDR-L1 tyrosine sulfated anti-hLAG3 antibodies and CDR-L1 tyrosine sulfated anti-hLAG3 antibodies are expressed from Chinese Hamster Ovary Cells, and wherein the purified composition is obtained by a method comprising the steps of:
- applying an aqueous mixture comprising CDR-L1 tyrosine sulfated and non-CDR-L1 tyrosine sulfated anti-hLAG3antibodies at a pH of about 6.5 to 7.0 to an anion exchange resin, and collecting a non-resin bound aqueous fraction of the aqueous mixture.
A purified composition comprising non-CDR-L1 tyrosine sulfated antibodies that specifically bind to human lymphocyte activation gene 3 (anti-hLAG3) comprising:
- a heavy chain amino acid sequence of SEQ ID NO:
- 52 and a light chain amino acid sequence of SEQ ID NO:
- 51 and less than about 0.5% CDR-L1 tyrosine sulfated anti-hLAG3antibodies as compared to non-CDR-L1 tyrosine sulfated anti-hLAG3 antibodies as determined by mass spectrometry, wherein the CDR-L1 tyrosine sulfated anti-hLAG3 antibodies comprise a heavy chain amino acid sequence of SEQ ID NO:
- 52 and a light chain amino acid sequence of SEQ ID NO:
- 51 with the tyrosine at CDR-L1 sulfated, wherein the non-CDR-L1 tyrosine sulfated anti-hLAG3 antibodies and the CDR-L1 tyrosine sulfated anti-hLAG3 antibodies are expressed from Chinese Hamster Ovary Cells, and wherein the purified composition is obtained by a method comprising the steps of:
- applying an aqueous mixture comprising CDR-L1 tyrosine sulfated and non-CDR-L1 tyrosine sulfated anti-hLAG3 antibodies at a pH of 6.5 to 7.5 to an anion exchange resin, and collecting a non-resin bound aqueous fraction of the aqueous mixture.
Dependencies are inferred from the claim text.