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US 12,600,761 B2Granted
Methods and compositions for purification of trimeric fusion proteins
AssigneeSichuan Clover Biopharmaceuticals, Inc.
InventorsPeng Liang, Joshua Liang
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Abstract
The present disclosure provides methods and compositions for affinity purification of proteins containing a C-terminal polypeptide sequence of a procollagen, such as trimeric fusion proteins. Also provided are polypeptides, fusion proteins, and recombinant polypeptides containing an Endo 180 ecto-domain or portion thereof capable of reversibly binding a C-terminal polypeptide sequence of a procollagen. Also provided are articles of manufacture, kits, and apparatus related to the Endo 180 ecto-domain polypeptides.
Claims17
A method for purifying a secreted trimeric fusion protein from a sample, comprising:
- (a) contacting a sample comprising the secreted trimeric fusion protein with a soluble Endo180 polypeptide linked to a support,
- wherein the trimeric fusion protein comprises a heterologous polypeptide sequence fused to a C-terminal polypeptide sequence of a procollagen, and
- wherein the soluble Endo180 polypeptide specifically binds to the C-terminal polypeptide sequence of the procollagen, whereby the secreted trimeric fusion protein and the soluble Endo180 polypeptide form a complex on the support;
- (b) removing unbound and/or nonspecifically bound molecules from the complex;
- and
- (c) dissociating the complex to release the trimeric fusion protein, thereby purifying the secreted trimeric fusion protein from the sample.
3 more elements
Dependencies are inferred from the claim text.