Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.2.1.23 (beta-galactosidase)
 14,648 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Clones have been isolated from the heterogeneous Spodoptera frugiperda IPLB-SF21-AE insect cell population. Five of these clones, in addition to the parent cell line and the SF9 cell line (another clonal isolate of the parent cell line), have been compared in regards to morphology, growth, budded virus synthesis, and recombinant protein synthesis. No significant differences in cell morphology were found among these cell lines. There was, however, a significant difference in the average cell size, with diameters ranging from 9.30 +/- 0.184 to 11.11 +/- 0.22 microns and from 9.17 +/- 0.05 to 11.25 +/- 0.24 microns for cells growing in Excell 401 serum-free medium in spinner flask cultures and in TNM-FH medium supplemented with 10% FBS in tissue flask cultures, respectively. While no significant differences in the growth rates were found in TNM-FH medium containing 10% calf serum, significant differences were found in Excell 401 serum-free medium, with population doubling times ranging from 38.5 +/- 6.6 to 64.5 +/- 6.4 h in spinner flask studies. Significant differences in expression levels of Escherichia coli beta-galactosidase (beta-gal) were also found in both 12-well plates and spinner flasks. In the 12-well plate studies, the peak levels of beta-galactosidase obtained by these cell lines ranged from 0.332 +/- 0.091 to 0.805 +/- 0.117 mg/10(6) cells and from 0.580 +/- 0.130 to 1.458 +/- 0.132 mg/10(6) cells in Excell 401 and Hyclone Hy-Q serum-free media, respectively. In the spinner flask studies, peak expression levels ranged from 0.128 +/- 0.053 to 0.573 +/- 0.215 mg/10(6) cells in Excell 401 serum-free medium.(ABSTRACT TRUNCATED AT 250 WORDS)
 ...
PMID:Clonal variation in the Spodoptera frugiperda IPLB-SF21-AE insect cell population. 776 39

Sf9 insect cells infected with a recombinant baculovirus expressing beta-galactosidase and suspended in fresh medium (TNM-FH supplemented with 10% fetal bovine serum) at the time of infection were cultured in shake flasks at various combinations of initial cell density and multiplicity of infection (MOI). The effects of cell density and MOI on beta-galactosidase production were quantitatively analyzed by plotting the beta-galactosidase yield against the time integral of the viable cell density from the time of infection to the time when the beta-galactosidase production reached a plateau. The beta-galactosidase yield had a maximum value at a viable cell density time integral of approximately 8 x 10(6) cells.d/cm3 for each MOI used in a range from 0.01 to 10 plaque-forming units per cell (pfu/cell). Since glucose and fructose were exhausted when the culture reached 8 x 10(6) cells.d/cm3, it was concluded that protein production in a high-cell-density culture was limited by nutrient depletion in the culture medium, and hence the nutritional capacity of the medium was able to be determined as the viable cell density time integral at which the maximum product yield was attained. In cultures infected at a low MOI (< or =1 pfu/cell), the specific productivity, and thereby the yield, of beta-galactosidase declined with decreasing MOI due to the reduction in the proportion of initially infected cells. These results indicate that production of a recombinant protein in a culture with medium replacement at the time of infection can be optimized if the cells are infected at a high MOI (> or = 1 pfu/cell) and at a cell density such that the viable cell density time integral reaches the nutritional capacity just as the protein production is completed.
 ...
PMID:Optimal production of recombinant protein by the baculovirus-insect cell system in shake-flask culture with medium replacement. 1623 31

The production of beta-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 beta-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 beta-galactosidase production, their removal by medium replacement on post-infection day 1 gave a beta-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 beta-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.
 ...
PMID:Recombinant protein production by the baculovirus-insect cell system in Basal media without serum supplementation. 1900 1