Solubility tags for the expression and purification of bioactive peptides
AssigneeE.I. du Pont de Nemours and Company
InventorsQiong Cheng, Linda Jane Decarolis, Stephen R. Fahnestock, Tanja Maria Gruber, Lisa Diane Reiss, Pierre E. Rouviere
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Abstract
Peptide tags, referred to here as inclusion body tags, are disclosed useful for the generation of insoluble fusion peptides. The fusion peptides comprise at least one inclusion body tag operably linked to a peptide of interest. Expression of the fusion peptide in a host cell results in a product that is insoluble and contained within inclusion bodies in the cell and/or cell lysate. The inclusion bodies may then be purified and the protein of interest may be isolated after cleavage from the inclusion body tag.
Claims14
An inclusion body tag comprising the structure:
- SEQ ID NO:
- 58 Spacer [[SEQ ID NO:
- 58]-[Spacer]m]n
- wherein
- SEQ ID NO:
- 58 is Gln-Gln-Xaa1-Phe-Xaa2-Trp-Xaa3-Phe-Xaa4-Xaa5-Gln;
- Xaa1=Arg, His, or Lys;
- Xaa2=Gln, His, or Lys;
- Xaa3=Gln, His, or Lys;
- Xaa4=Glu or Gln;
- Xaa5=Gln or Lys;
- n=1 to 10;
- m=n−1;
- and
- Spacer=is a peptide comprising the amino acids selected from the group consisting of proline, arginine, glycine, glutamic acid, and cysteine.
Dependencies are inferred from the claim text.