NDLI: Improvement of the yields of recombinant actin and myosin V–HMM in the insect cell/baculovirus system by the addition of nutrients to the high-density cell culture

Content Provider	Springer Nature Link
Author	Ohki, Takashi Mikhailenko, Sergey V. Arai, Tomomi Ishii, Shuya Ishiwata, Shin’ichi
Copyright Year	2012
Abstract	Baculovirus infection of Sf9 cells at high densities, such as during mid- and late exponential phase, often results in a significant reduction of protein yield per cell, compared to the early exponential phase. Nutrient depletion has been considered as a major cause for the decreased protein yield. In this study, we report that the addition of nutrients (glucose, yeastolate ultrafiltrate, and lactalbumin hydrolysate) and small fraction of fresh medium at time of infection restores the expression level of actin and myosin V–HMM at late exponential phase (11.3 × 10$^{6}$ cells/ml) to that at early exponential phase (1.0 × 10$^{6}$ cells/ml). The relative yields of actin and myosin V–HMM were approximately equal at both phases (typically 200 mg of actin and 5 mg of myosin V–HMM per 10$^{10}$ cells), i.e., the volumetric yield of proteins from the cell culture at late exponential phase was approximately tenfold higher than at early exponential phase. The functionality of the recombinant actin and myosin V–HMM was confirmed by measuring the rate of actin polymerization, actin-activated ATPase, and the gliding velocity of actin filaments in an in vitro motility assay.
Starting Page	351
Ending Page	358
Page Count	8
File Format	PDF
ISSN	01424319
Journal	Journal of Muscle Research & Cell Motility
Volume Number	33
Issue Number	5
e-ISSN	15732657
Language	English
Publisher	Springer Netherlands
Publisher Date	2012-09-19
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Actin Baculovirus Myosin V Insect cell High-density culture Nutrient feeding Proteomics Biomedicine general Cell Biology Animal Anatomy / Morphology / Histology
Content Type	Text
Resource Type	Article
Subject	Cell Biology Physiology Biochemistry

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