Soluble Intein Fusion Proteins And Methods For Purifying Biomolecules
AssigneeMerck Patent GmbH
InventorsMartin Zillmann, Joe Orlando
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Abstract
The present invention relates to fusion proteins comprising an N-intein polypeptide and an N-intein solubilization partner, and affinity chromatography matrices comprising such fusion proteins, as well as methods of using same.
Claims28
A soluble fusion protein comprising an N-intein polypeptide capable of forming an active intein complex by associating with a second fusion protein comprising a C-intein polypeptide, wherein the N-intein polypeptide can be covalently attached to a solid support.
A matrix comprising an N-intein polypeptide attached, optionally by covalent attachment, to a solid support.
A split intein-based affinity chromatography system, comprising:
- a) a first fusion protein comprising a C-intein polypeptide joined to a target molecule by a peptide bond;
- and
- b) an affinity chromatography matrix comprising an N-intein polypeptide that is attached, optionally by covalent attachment, to a solid support.
An expression vector comprising a nucleic acid encoding a fusion protein comprising a C-intein polypeptide joined to a target molecule by a peptide bond.
A nucleic acid encoding a fusion protein comprising an N-intein polypeptide and an N-intein solubilization partner joined by a peptide bond, wherein the N-intein solubilization partner has a molecular weight of less than about 15 kDa, an Aliphatic Index value less than about 60, and a Grand Average Hydropathy value less than −1, and enhances solubility of the N-intein polypeptide compared to the N-intein polypeptide expressed in the absence of the solubilization partner.
Dependencies are inferred from the claim text.