| abstract |
The invention relates to a method for producing any protein in a form that can be quickly isolated from solution using a specific monoclonal antibody, "HPC-4". It has now been determined that it is possible to form a fusion protein composed of the epitope and a protein to be isolated, and to isolate the protein via HPC-based affinity chromatography. -4. According to a preferred embodiment, a specific segmentation site by protease is inserted between the epitope and the protein so as to be able to easily remove the epitope from the isolated protein. According to one example, a soluble tissue factor with functional activity, and comprising the epitope of twelve amino acids recognized, in combination with calcium, by HPC-4, as well as a site of segmentation by factor Xa, was expressed from a vector introduced into the prokaryotic expression system. Recombinant tissue factor can be quickly isolated in a single chromatographic step using HPC-4 monoclonal antibody immobilized on a suitable substrate. Once the protein is isolated, the protein C epitope is removed by factor Xa segmentation, thus leaving only the soluble tissue factor in functional activity. |