Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues
Patent
1992-01-03
1994-03-29
Hill, Jr., Robert J.
Chemistry: natural resins or derivatives; peptides or proteins;
Proteins, i.e., more than 100 amino acid residues
435 681, 435 697, 435 696, 530381, 530413, 53038825, 530413, C07K 1300
Patent
active
052985997
ABSTRACT:
A method is disclosed to make any protein in a form that can be isolated rapidly from a solution using a specific monoclonal antibody designated "HPC-4". It has now been determined that it is possible to form a fusion protein of the epitope with a protein to be isolated, and isolate the protein using HPC-4-based affinity chromatography. In the preferred embodiment, a specific protease cleavage site is inserted between the epitope and the protein so that the epitope can be easily removed from the isolated protein. In an example, a functionally active soluble tissue factor including the twelve amino acid epitope recognized in combination with calcium by HPC-4 and a factor Xa cleavage site was expressed from a vector inserted into a procaryotic expression system. The recombinant tissue factor can be rapidly isolated in a single chromatographic step using the HPC-4 monoclonal antibody immobilized on a suitable substrate. Once isolated, the Protein C epitope is removed by cleavage with factor Xa, leaving the functionally active, soluble tissue factor.
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United States government has rights in this invention by virtue of grants from the National Institutes of Health, grant numbers R01 HL44225 and R01 HL29807.
Esmon Charles T.
Morrissey James H.
Rezaie Alireza
Fitzgerald David L.
Hill Jr. Robert J.
Oklahoma Medical Research Foundation
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