Large-scale purification and characterization of recombinant tick anticoagulant peptide |
| |
Authors: | E Dale Lehman Ted F Schaefer Craig T Przysiecki Joseph G Joyce F James Bailey Cheryl A Schulman Carl J Burke Harri G Ramjit William J Miller |
| |
Institution: | E. Dale Lehman*, Ted F. Schaefer, Craig T. Przysiecki, Joseph G. Joyce, F. James Bailey, Cheryl A. Schulman, Carl J. Burke, Harri G. Ramjit,William J. Miller |
| |
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. |
| |
Keywords: | |
本文献已被 ScienceDirect 等数据库收录! |
|