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| LARGE-SCALE PURIFICATION AND CHARACTERIZATION OF RECOMBINANT TICK ANTICOAGULANT PEPTIDE |
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| Author(s): LEHMAN ED, SCHAEFER TF, PRZYSIECKI CT, JOYCE JG, BAILEY FJ, SCHULMAN CA, BURKE CJ, RAMJIT HG, MILLER WJ |
| Source: JOURNAL OF CHROMATOGRAPHY-BIOMEDICAL APPLICATIONS Volume: 574 Issue: 2 Pages: 225-235 Published: FEB 14 1992 |
| Times Cited: 16 References: 26 |
| Abstract: Recombinant tick anticoagulant peptide (r-TAP), a potent and specific inhibitor of blood coagulation factor Xa, was purified to > 99% homogeneity at the multi-gram scale. Genetically engineered yeast secreted 200-250 mg/l of the heterologous protein into the medium. Cells were separated from broth by diafiltration and purification was done by two chromatographic steps, both conducive to operation on a large scale. Analysis of the purified protein by several methods indicated that it was > 99% homogeneous and no incompletely processed or truncated proteins were detected. Physico-chemical characterization data of r-TAP show that it exists as a monomer in solution and no evidence of post-translational modification was observed. The purified protein was fully active in inhibiting human coagulation factor Xa. |
| Document Type: Article |
| Language: English |
| Reprint Address: LEHMAN, ED (reprint author), MERCK SHARP & DOHME LTD, DEPT CELLULAR & MOLEC BIOL, W POINT, PA 19486 USA |
Addresses:
1. MERCK SHARP & DOHME LTD, DEPT PHARMACEUT RES & DEV, W POINT, PA 19486 USA
2. MERCK SHARP & DOHME LTD, DEPT DRUG METAB, W POINT, PA 19486 USA |
| Publisher: ELSEVIER SCIENCE BV, PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS |
| Subject Category: Chemistry, Analytical |
| IDS Number: HG322 |
| ISSN: 0378-4347 |
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| | | | | | | | | Record from Web of Science® | | | | | | | | | |