Monolithic peptidyl sorbents for comparison of affinity properties of plasminogen activators.
Plasminogen activators are the proteases which convert plasminogen into plasmin dissolving, in its turn, the major component of blood clots, fibrin. They are extremely useful in heart attack therapy. Modern and most appropriate way of scaled up production of these valuable proteins is gene engineering. In this case, a separation and a purification of target product become the important steps of the whole process. Recently developed affinity chromatography on short monolithic columns seems to be a very attractive method for these purposes. High speed of a process prevents the protein's denaturation due to temperature or/and solvents influence. The better mass transfer mechanism (convection rather than diffusion) allows considering only biospecific complexing as time limiting step. Specificity of several synthetic peptides to plasminogen activators have been studied by affinity chromatography on short monolithic columns. Peptide ligands were synthesized by conventional solid phase peptide synthesis (SPPS). The immobilization procedure was carried out as a one step process at static conditions. The results of quantitative evaluation of such affinity interactions were compared with those established for plasminogen that is the natural affinity counterpart to both proteases. Additionally, some of investigated peptides were synthesized directly on GMA-EDMA disks and their affinity properties were compared with those established for the case of immobilized ligands. The possibility of using of synthetic peptidyl ligands for plasminogen activators isolation from native cell supernatant and model protein mixtures has been demonstrated.
Vlakh
Evgenia G
EG
Institute of Macromolecular Compounds, Russian Academy of Sciences, Bolshoy Pr. 31, 199 004 St. Petersburg, Russia.
Tappe
Alexander
A
Kasper
Cornelia
C
Tennikova
Tatiana B
TB
D000327:Adsorption; D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002846:Chromatography, Affinity; D006224:Cricetinae; D003432:Cross-Linking Reagents / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D007202:Indicators and Reagents; D008689:Methacrylates / Q000737:chemistry; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry*; D010960:Plasminogen Activators / Q000737:chemistry*; D011786:Quality Control; D013056:Spectrophotometry, Ultraviolet
D003432:Cross-Linking Reagents; D007202:Indicators and Reagents; D008689:Methacrylates; D010455:Peptides; C004919:ethylene dimethacrylate; D010960:Plasminogen Activators