Method of protein purification

摘要

A method for producing a target protein is provided, which includes steps described below. A crude extract including a fusion protein is provided. The fusion protein includes a tag, a target protein and a linker inserted between the tag and the target protein. The fusion protein and magnetic particles are then bound to form a magnetic particle-binding fusion protein. Finally, the linker of the magnetic particle-binding fusion protein undergoes autocleavage by using a cleavage buffer solution to release the target protein. A one-pot process for producing a purified target protein is also provided.

主权项

1. A method for producing a target protein, comprising steps of:
(a) providing a crude extract including a fusion protein, the fusion protein comprising a tag, a target protein and a linker inserted between the tag and the target protein, wherein the linker has an amino acid sequence of SEQ ID NO:1; (b) binding magnetic particles and the fusion protein of the crude extract to form a magnetic particle-binding fusion protein; and (c) enabling autocleavage of the linker of the magnetic particle-binding fusion protein by using a cleavage buffer solution to release the target protein, wherein the cleavage buffer is TRIS buffer or phosphate buffer.