Large-scale production and purification of recombinant protein from an insect cell/baculovirus system in Erlenmeyer flasks: application to the chicken poly(ADP-ribose) polymerase catalytic domain.
A simple and inexpensive shaker/Erlenmeyer flask system for large-scale cultivation of insect cells is described and compared to a commercial spinner system. On the basis of maximum cell density, average population doubling time and overproduction of recombinant protein, a better result was obtained with a simpler and less expensive bioreactor consisting of Erlenmeyer flasks and an ordinary shaker waterbath. Routinely, about 90 mg of pure poly(ADP-ribose) polymerase catalytic domain was obtained for a total of 3 x 10(9) infected cells in three liters of culture.
Miranda
E A
EA
Ecole Supérieure de Biotechnologie de Strasbourg, UPR 9003 du CNRS, France. everson@feq.unicamp.br
de-Murcia
G
G
Ménissier-de-Murcia
J
J
D000818:Animals; D016367:Baculoviridae; D066298:In Vitro Techniques; D007313:Insecta / Q000166:cytology*; D011064:Poly Adenosine Diphosphate Ribose; D011062:Polynucleotide Adenylyltransferase / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000302:isolation & purification*
D011994:Recombinant Proteins; D011064:Poly Adenosine Diphosphate Ribose; D011062:Polynucleotide Adenylyltransferase