Successful protocols have been designed for automated transient transfection of human embryonic kidney (HEK) 293T and 293S GnTI− cells in various flask formats.
The fluid shear stress, which was generated by various flask shaking intensities, could reduce the mass transfer resistance in the periplasmic layer [23],[23].
Briefly, 50 mL of 0.01 M KNO3 solutions were placed in various Erlenmeyer flasks.
The CDMSS allows the determination of the volumetric oxygen transfer coefficients of various types of flasks and of gas transfer resistance through various types of culture plugs.
As the field has developed, kL a values have increased to >360 h-1 for microbioreactors (Funke et al., 2010a; Lee et al., 2006) and >1,000 h-1 for minibioreactors (Puskeiler et al., 2005), allowing analogy with culture systems increasing in scale from microwell plates, through shaker flasks, various stirred tank reactors (STRs), and up to production scale bioreactors.
For gaining a better understanding and control of shake flask cultivations, various methods for online monitoring of process parameters in shake flask cultivations have been developed in recent years.
Long term decoloration experiments were initiated by adding 5 mL wet volume of fungal biomass from 5 day old culture in liquid Kirk's medium without amaranth to 100 mL culture flasks containing various media.
The same protocol was used for feeding Williams 82 seedlings with INA for various hours in Erlenmeyer flasks.
In a first series of shake-flask experiments, various media were prepared like the just-described non-supplemented DW medium with the modification that various sources of nitrogen (10 g/L (NH4 2SO4, 10 g/L (NH4 2CO3, or 5 g/L urea) were added before medium sterilization.
To demonstrate and compare the efficiencies of SsAraT, AtStp2 and ScGal2 in L-arabinose utilization by recombinant S. cerevisiae cells, growth and sugar consumption were characterized in strain TSY01 expressing the various transporters by shake-flask aerobic batch cultivations.
