Insect cells quantification

Riley MR, Arnold MA, Murhammer DW, Walls EL, DelaCruz N. Adaptive calibration scheme for quantification of nutrients and byproducts in insect cell bioreactors by near-infrared spectroscopy. Biotechnology Progress 1998, 14, 527-533. 

The insect cells were seeded in 2.5 ml of complete TClOO medium as described above. After the cells had attached, a viral inoculum (200-400 jj,l) was added to the cells. At the same time, a mixture of [ Sjmethionine and cysteine (0.5 mCi per 25-cm culture flask) was added to the medium. The time course of intracellular radioactive incorporation into total protein was analyzed over 72 hr. The cells were harvested and washed three times with cold PBS. The final cell pellet was resuspended in 300 /xl of lysis buffer, boiled for 5 min, and stored frozen at — 20°C for later analysis. Proteins were analyzed on 10% SDS-PAGE using 5-10 /jlI of the lysate. Radioactive incorporation into protein was detected after fluorography using EN HANCE (DuPont de Nemours) according to the manufacturer s protocol. Exposure of the dried gel on X-ray film was followed by laser densitomet-ric quantification. In parallel, after electrophoretic separation of the proteins, the gel was stained with Coomassie Brilliant Blue and dried, and the respective recombinant proteins were then cut out with a razor blade. Specific radioactive incorporation was measured by liquid scintillation counting. 

A. Expression and Quantification of Radiolabeled Recombinant sGC in HighFive Insect Cells... 

Trinh K, Garcia-Briones MA, Hink FH, Chalmers JJ. (1994) Quantification of damage to suspended insect cells as a result of bubble rupture. Biotechnol. Bioeng., 43 37-45. VaUez-chetreanu F. (2006) Characterization of the mechanism of action of spin-filters for animal cell perfusion cultures. These no 3488 (2006), Ecole Polytechnique Eederale de Lausanne, Lausanne, France. 

No less revolutionary in the quantification of pharmacodynamics was the insistance on size relationships which Alfred Joseph Clark (London) introduced in his book, The Mode of Action of Drugs on Cells (Clark, 1933). The mnemonic diagram that forms the frontispiece of the present book will serve to introduce this aspect. In it we see a typical mammal (dog) followed by a typical insect (flea) which is a thousand times smaller, and this is followed by a typical microbe (streptococcus) which is yet a thousand times smaller, and finally a typical bioactive molecule (p-aminobenzoic acid) which is one thousand times smaller still. Thus we have familiar reference objects staked out over a size-range of 10 (a million-millionfold). 

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