Production and purification of a protein fused to a binding...

Details Production and purification of a protein fused to a binding... Production and purification of a protein fused to a binding...

C - Chemistry, Metallurgy

12

N

195/1.22, 195/1.

C12N 15/62 (2006.01) C07K 1/22 (2006.01) C07K 19/00 (2006.01) C12N 15/52 (2006.01)

Patent

CA 1340173

Methods and products are provided for producing and/or purifying virtually any hybrid polypeptide molecule employing recombinant DNA techniques. More specifically, a DNA fragment coding for a protein molecule, e.g. a polypeptide or portion thereof, is fused to a DNA fragment coding for a binding protein such as the gene coding for the maltose binding protein. The fused DNA is inserted into a cloning vector and an appropriate host transformed. Upon expression, a hybrid polypeptide is produced which can be purified by contacting the hybrid polypeptide with a ligand or substrate to which the binding protein has specific affinity, e.g. by affinity chromatography. The hybrid polypeptide so purified may in certain instances be useful in its hybrid form, or it may be cleaved to obtain the protein molecule itself by, for example, linking the DNA fragments coding for the target and binding proteins with a DNA segment which codes for a peptide which is recognized and cut by a proteolytic enzyme. The present invention also relates to certain vectors useful in practing the above process as well as to a bioreactor and methods employing the bound hybrid polypeptide, e.g. where the bound target polypeptide is contacted and reacted with a sustance which interacts with the bound target polypeptide to produce a desired result.

560985

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