Methods for production of receptor and ligand isoforms
OwnerPei Jin, H. Shepard +2inventors
InventorsPei Jin, H. Shepard, Cornelia Gorman, Juan Zhang
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Abstract
Provided are methods for production of cell surface receptor (CSR) and ligand isoforms. In particular, isoform fusions that a precursor sequence for secretion, processing and intracellular trafficking are provided. Nucleic acid molecules encoding the fusions are expressed in a host cell and the encoded and partially or completely processed encoded CSR or ligand isoforms is produced in the cell culture medium. The resulting polypeptide optionally includes an epitope tag for the detection and/or purification thereof.
Claims114
A polypeptide, comprising a receptor tyrosine kinase (RTK) isoform operatively linked directly or indirectly via a polypeptide linker to a heterologous precursor sequence or a sufficient portion thereof to effect secretion, processing and/or trafficking of the linked RTK intron fusion protein.
A polypeptide, comprising a Receptor for Advanced Glycation Endproducts (RAGE) isoform operatively linked directly or indirectly via a polypeptide linker to a heterologous precursor sequence or a sufficient portion thereof to effect secretion and/or trafficking of the RAGE isoform, wherein the polypeptide optionally includes a tag that facilitates polypeptide purification and/or detection.
A polypeptide, comprising a tumor necrosis factor receptor (TNFR) isoform operatively linked directly or indirectly via a linker to a heterologous precursor sequence or a sufficient portion thereof to effect secretion, processing and/or trafficking of the TNFR isoform, wherein the polypeptide optionally includes a tag that facilitates polypeptide purification and/or detection.
A nucleic acid molecule, comprising a sequence of nucleotides set forth in any one of SEQ ID NOS:
- 31, 33, 35, 37, 39, 41, 43, 45 and 47.
A method of producing a CSR isoform or a ligand isoform, comprising introducing a nucleic acid molecule encoding the isoform and a signal sequence, whereby the isoform is secreted from the cell.
A polypeptide, comprising a cell surface receptor (CSR) or ligand isoform wherein:
- the polypeptide lacks an endogenous precursor sequence;
- and
- the polypeptide contains one or more additional amino acids at its N-terminus.
A polypeptide, comprising a hepatocyte growth factor (HGF) isoform operatively linked directly or indirectly to a heterologous precursor sequence or a sufficient portion thereof to effect secretion and/or trafficking of the HGF isoform, wherein the polypeptide optionally includes a tag that facilitates polypeptide purification and/or detection.
Dependencies are inferred from the claim text.