Production of vampire bat plasminogen activator DSPA alpha 1 in CHO and insect cells.
Salivary plasminogen activator from the vampire bat Desmodus rotundus (DSPA alpha 1) is a promising new thrombolytic agent. Continuous growth of a stably transfected, methotrexate amplified, dhfr- CHO cell line yields up to 60 mg l-1 of DSPA alpha 1. Utilizing an engineered baculovirus 10 mg l-1 were produced in batches of Sf9 insect cells. Recombinant DSPA alpha 1 is purified from both sources using a one-step purification protocol. Although differences in glycosylation were detected, enzymatic activity and fibrin cofactor dependency are unaffected when DSPA alpha 1 derived from the two expression systems is compared.
Petri
T
T
Research Laboratories of Schering AG, Berlin, Germany.
Langer
G
G
Bringmann
P
P
Cashion
L
L
Shallow
S
S
Schleuning
W D
WD
Donner
P
P
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001709:Biotechnology; D016466:CHO Cells; D002460:Cell Line; D002685:Chiroptera; D006224:Cricetinae; D005343:Fibrinolytic Agents / Q000302:isolation & purification; D005784:Gene Amplification; D006031:Glycosylation; D010957:Plasmids / Q000235:genetics; D010960:Plasminogen Activators / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D018411:Spodoptera; D014162:Transfection
D005343:Fibrinolytic Agents; D011994:Recombinant Proteins; C069588:salivary plasminogen activator alpha 1, Desmodus rotundus; D010960:Plasminogen Activators