Abstract
We have investigated large scale production processes (up to 2 liters) of recombinant proteins using the baculovirus expression system in order to optimize the product yields. Experiments using cell lines ofSpodoptera frugiperda (Sf9) andMamestra brassicae (IZD-Mb0503) were performed to show the different production capacities of the cell lines. The influence of the infection at different cell densities is described. Beyond that, TC100-, IPL41- and serum-free IPL41-medium were compared to demonstrate their different capabilities of supporting cell growth and protein expression. Additionally, the inhibitory effect of FCS on the protease activity of kallikrein, which is produced in its zymogenic form, is discussed. Improved production parameters are described, which enabled us to produce up to 8000 units of activated pro-kallikrein within 14 days using perfusion cultivation.
Similar content being viewed by others
Abbreviations
- AcNPV:
-
autographa californica nuclear polyhedrosis virus
- MOI:
-
multiplicity of infection
References
Adamson SR, Fitzpatrick SL and Behie LA (1983)In vitro production of high titre monoclonal antibody by hybridoma cells in dialysis culture. Biotechnol. Lett. 5:573–576.
Angermann A, Rahn HP, Hektor T, Fertig G and Kemme M (1992) Purification and characterization of human salivary-gland prokallikrein from recombinant baculovirus-infected insect cells. Eur. J. Biochem. 206:225–233.
Bailey GS (1988) Rat pancreas kallikrein. Meth. Enzymol. 163: 115–129.
De Gooijer CD, Van Lier FLJ, Van Den End EJ, Vlak JM and Tramper J (1989) A model for baculovirus production with continuous insect cell cultures. Appl. Microbiol. Biotechnol. 30:497–501.
Fertig G and Klöppinger M (1991) Cell cycle kinetics of insect cells and the application in fermentation technology. In Vitro 27:105A.
Fiedler F, Geiger R, Hischauer C and Leysath G (1978) Peptide esters and nitroanilides as substrates for the assay of human urinary kallikrein. Hoppe-Seyler's Z. Physiol. Chem. 259: 1667–1673.
Fraser MJ (1989) Expression of eukaryotic genes in insect cell cultures. In Vitro Cell. Develop. Biol. 25:225–235.
Fung MC, Chin KYM, Weber T, Chang TW and Chang NT (1988) Detection and purification of a recombinant human B lymphotropic virus (HHV-6) in the baculovirus expression system by limiting dilution and DNA dot-blot hybridization. J. Virologic. Meth. 19:33–42.
Gardiner GR and Stockdale H (1975) Two tissue culture media for production of lepidopteran cells and nuclear polyhedrosis viruses. J. Invert. Path. 25:383–370.
Harlow E and Lane D (1988) Antibodies. A Laboratory Manual. Cold Spring Harbor, New York.
Hink WF, Strauss EM and Ramoske WA (1977) Propagation of Autographa californica nuclear polyhedrosis virus in cell culture: methods for infecting cells. J. Invertebr. Pathol. 30: 185–191.
Hink WF, Thomsen DR, Davidson DJ, Meyer AL and Castellino FJ (1991) Expression of three recombinant proteins using baculovirus vectors in 23 insect cell lines. Biotechnol. Prog. 7:9–14.
Klöppinger M, Fertig G, Fraune E and Miltenburger HG (1990). Multistage production of Autographa californica nuclear polyhedrosis virus in insect cell cultures. Cytotechnology 4: 271–278.
Kuroda K, Hauser C, Rott R, Klenk ND and Doerfler W (1986) Expression of the influenza virus haemagglutinin in insect cells by a baculovirus vector. EMBO J. 5:1359–1365.
Lehmann J, Vorlop J and Büntemeyer H (1988) Bubble-free reactors and their development for continuous culture with cell recycle. In: Spier RE and Griffiths JB (eds) Animal Cell Biotechnology, pp. 221–237, Academic Press Ltd., London.
Luckow VA and Summers MD (1988) Trends in development of baculovirus expression vectors. Bio/Technology 6:47–55.
Lynn DE and Hink WF (1978) Infection of synchronized TN-368 cell cultures with alfalfa looper nuclear polyhedrosis virus. J. Invertebr. Pathol. 32:1–5.
Maiorella B, Inlow D, Shauger A and Harano D (1988). Largescale insect cell-culture for recombinant protein production. Bio/Technology 6:1406–1410.
Maniatis T, Fritsch EF and Sambrock J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.
Miller LK (1988) Baculoviruses as gene expression vectors. Ann. Rev. Microbiol. 42:177–199.
Miltenburger HG, David P, Mahr U and Zipp W (1977) Establishment of lepidopteran cell lines andin vitro replication of insect pathogenic viruses. I.Mamestra brassicae cell lines and NPV replication. Z. Angew. Entomol. 82:306–323.
Miltenburger HG (1983) Investigations on the cultivation of insect cell lines in serum-free media. In: Fischer G and Wieser RJ (eds) Hormonally-Defined Medium, pp. 31–45. Springer Verlag, Berlin.
Müller-Esterl W, Iwanaga S and Nakanishi S (1986) Kininogens revisited. Biochem. Sci. 11:336–339.
Murhammer DW and Goochee CF (1988) Scaleup of insect cell cultures: protective effects of pluronic F-68. Bio/Technology 6:1411–1418.
Regoli D and Barabe J (1980) Pharmacology of bradykinin and related kinins. Pharmacol. Rev. 32:1–46.
Schill WB, Preissler G, Dittmann B and Müller-Esterl W (1979) Effect of pancreatic kallikrein, spermacrosin and HMW kininogen on cervical mucus penetration ability of seminal plasma-free human spermatozoa. Adv. Exp. Med. Biol. 120: 305–310.
Summers MD and Smith GD (1987) A Manual of Methods for Baculovirus and Insect Cell Culture Procedures. Texas Agricultural Experiment Station, Bulletin, No 1555.
Takada Y, Skidgel RA and Erdös EG (1985) Purification of human urinary prokallikrein. Biochem. J. 232:851–858.
Tramper J, Van Den End EJ, De Gooijer CD, Kompier R, Van Lier FLJ, Usmany M and Vlak JM (1990) Production of baculovirus in a continuous insect-cell culture. Annals of the New York academy of sciences, biochemical engineering 289: 423–430.
Van Lier FLJ, Van Den End EJ, De Gooijer CD, Vlak JM and Tramper J (1990) Continuous production of baculovirus in a cascade of insect-cell reactors. Appl. Microbiol. Biotechnol. 33:43–47.
Weiss SA, Smith GC, Kalter SS and Vaughn JL (1981) Improved method for the production of insect cell cultures in large volume. In Vitro 17:495–502.
Weiss SA, DiSorbo DM, Whitford WG and Godwin G (1991) Improved production of recombinant proteins in high density insect cell culture. In Vitro 27:42A.
Wicklmayer M, Dietze G, Mayer L, Böttger T and Gunst J (1979) Evidence for an involvement of kinin liberation in the priming action of insulin on glucose uptake into sceletal muscle. FEBS Lett. 98:61–65.
Wood HA, Johnston LB and Burand JP (1982) Inhibition of Autographa californica nuclear polyhedrosis virus replication in high-density Trichoplusia ni cell cultures. Virology 119: 245–251.
Youngblood WW and Kizer JS (1983) Strategies for the preparation of haptens for conjugation and substrates for iodination for use in radioimmunoassays of small oligopeptides. Meth. Enzymol. 103:407–431.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Fertig, G., Rahn, H.P., Angermann, A. et al. Biotechnological aspects of the production of human pro-kallikrein using the AcNPV-baculovirus-expression system. Cytotechnology 11, 67–75 (1993). https://doi.org/10.1007/BF00749059
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00749059