Large-Scale Insect Cell-Culture for Recombinant Protein Production

Maiorella, B.; Inlow, Duane; Shauger, Andrea; Harano, David

doi:10.1038/nbt1288-1406

Cited by 267 publications

(137 citation statements)

References 27 publications

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“…In the case of serum-free TNM-FH (Figure 1b), cells grew at a slower rate and reached 6 Â 10 5 cells/cm 3 , or 22% of the maximum cell density obtained in the serum-containing medium. Admixture of lipids, which are reported to stimulate serum-free growth of insect cells (Maiorella et al 1988), did not improve cell growth in Grace's medium, but did in TNM-FH. Although PA has been shown to promote the growth of Chinese hamster ovary (CHO) cells in serum-free culture (Sakai et al 1999;2002), supplementation of Grace's medium and TNM-FH with PA from egg yolk lecithin was not effective in increasing the density of Sf9 cells.…”

Section: Cell Growth In Basal Mediamentioning

confidence: 76%

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Recombinant Protein Production by the Baculovirus–Insect Cell System in Basal Media Without Serum Supplementation

Nishikawa¹,

Yamaji²,

Fukuda³

2003

Cytotechnology

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The production of b-galactosidase by Sf9 cells infected with recombinant Autographa californica nucleopolyhedrovirus (AcNPV) was investigated in shake-flask culture using two serum-free basal media: Grace's medium and TNM-FH (Grace's medium supplemented with lactalbumin hydrolysate and yeast extract). At the time of infection, cells grown in serum-supplemented TNM-FH were transferred into fresh basal media without adaptation. The absence of serum depressed the b-galactosidase yield considerably in Grace's medium, but to a much lesser extent in TNM-FH, where it reached around 2/3 of the level obtained in TNM-FH supplemented with 10% fetal bovine serum (FBS). While both lactalbumin hydrolysate and yeast extract promoted b-galactosidase production, their removal by medium replacement on post-infection day 1 gave a b-galactosidase yield nearly equal to that obtained in their continuous presence. Supplementation of basal media with phosphatidic acid (PA) from egg yolk lecithin, which has been shown to enhance cell growth and recombinant protein production in serum-free culture of Chinese hamster ovary (CHO) cells, was also effective in increasing b-galactosidase yield. Elevating the multiplicity of infection (MOI) from 2 to 10 plaque-forming units per cell (pfu/cell) also resulted in an increase in product yield. These results provide information important to the development of cost-effective serum-free culture technology for use in large-scale production of recombinant proteins by the baculovirus-insect cell system.

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Section: Cell Growth In Basal Mediamentioning

confidence: 76%

“…These problems have led to the development of serumfree media for use in insect cell culture. Serum-free media are currently available commercially, but more cost-effective medium formulations are still required for large-scale production of recombinant proteins (Maiorella et al 1988;Jesionowski and Ataai 1997;Donaldson and Shuler 1998).…”

Section: Introductionmentioning

confidence: 99%

Recombinant Protein Production by the Baculovirus–Insect Cell System in Basal Media Without Serum Supplementation

Nishikawa¹,

Yamaji²,

Fukuda³

2003

Cytotechnology

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“…Larder, personal communication). The baculovirus system has enabled us to obtain very efficient expression of both HCMV and VZV DNA polymerases, has given similar success with the HSV enzyme and could allow large scale production of these enzymes (Maiorella et al, 1988). There are still considerable problems with the system in terms of the quantity of the active enzyme that we can obtain in a soluble form, but the VZV-and HCMV-infected cell systems yield far smaller amounts.…”

mentioning

confidence: 99%

High level expression of DNA polymerases from herpesviruses

Ertl

Thomas²,

Powell³

1991

Journal of General Virology

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The DNA polymerase genes of human cytomegalovirus (HCMV) and varicella-zoster virus (VZV) were inserted separately into the polyhedrin gene of Autographa ealifornica nuclear polyhedrosis virus (AcNPV) by cotransfection of Spodoptera frugiperda (SFg) cells with baculovirus transfer vectors carrying the genes and AcNPV infectious DNA. Infection of SF9 cells with the recombinant viruses resulted in expression from the polyhedrin promoter of proteins of the expected Mrs. These proteins possessed DNA polymerase activities similar to that of the enzymes induced by the respective herpesvirus in infected cells, and were identified as HCMV and VZV DNA polymerase using inhibitors and specific antisera reactive with each enzyme.

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“…The formulation contained a basal medium of IPL-41 without carbohydrates and was supplemented with 4.0 g/L yeastolate (Difco) ultrafiltrate (10,000 dalton cutoff), 2.5 g/L tryptose phosphate broth (TPB) (Difco), 2.0 g/L Pluronic-F68 (BASF), 10.0 mL/L lipid concentrate (Life Technologies, Grand Island, NY), and 5.0 g/L glucose. The osmolality of the medium was adjusted to between 360 and 380 mOsm using NaCI.…”

Section: Serum-free Medium (Icsf-wb)mentioning

confidence: 99%

Production of recombinant proteins in high‐density insect cell cultures

Reuveny

Kim

Kemp

et al. 1993

Biotech & Bioengineering

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The effect of the growth phase of Spodoptera frugiperda (Sf9) cells on the production of recombinant proteins (β‐galactosidase and glucocerebrosidase) was investigated. Cells infected with the recombinant Autographa californica nuclear polyhedrosis virus at the late exponential and stationary phases yielded low quantities of expressed protein. Highest enzyme yields were obtained using Sf9 cells from the early exponential phase (0.9 mg β‐galactosidase/106 cells and 1.7 μg glucocerebrosidase/106 cells). Infection of resuspension of cells collected from various phases of growth in fresh medium resulted in 75% restoration of maximal expression levels. This finding suggested either nutrient limitation or waste product accumulation as the cause of the decrease in productivity at the latter phases of growth. Further experiments revealed that the highest productivity levels could be obtained with cultures of Sf9 cells grown in a fermentor to a cell concentration of 4 × 106 mL−1. The medium needed to be replaced prior to infection with the recombinant virus and supplemented with a mixture of glucose, L‐glutamine, and yeastolate ultrafiltrate. © 1993 John Wiley & Sons, Inc.

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Large-Scale Insect Cell-Culture for Recombinant Protein Production

Cited by 267 publications

References 27 publications

Recombinant Protein Production by the Baculovirus–Insect Cell System in Basal Media Without Serum Supplementation

Recombinant Protein Production by the Baculovirus–Insect Cell System in Basal Media Without Serum Supplementation

High level expression of DNA polymerases from herpesviruses

Production of recombinant proteins in high‐density insect cell cultures

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