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We successfully transduced Fabry patient CD34+ hematopoietic cells with "near-clinical grade" LV/AGA in small-scale cultures and then validated a clinically directed scale-up transduction process in a GMP-compliant cell processing facility.
Though some mutations clearly dampened protein expression, deletion of amino acids 57 70, corresponding to the predicted 3rd alpha helix of α-mating factor secretion signal, increased secretion of reporter proteins horseradish peroxidase and lipase at least 50% in small-scale cultures.
These small-scale cultures were compared for levels of expression of the GST fusion protein by anti-GST Western blot.
This will lead to balanced pH-regulation in small-scale cultures in the future.
Small-scale cultures were illuminated by 250 μmol photons s-1 m-2.
Small-scale cultures were inoculated to an OD730 of 0.2 to 0.3 and large-scale cultures were inoculated to an OD730 of approximately 0.1.
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However, at present, only small-scale culturing of a limited number of species is possible (Castellanos-Martínez and Gestal 2013).
The use of these systems allow to synthetize up to 28 mg/l of naringenin in small-scale culture.
This set-up is widely used in the early stages of small-scale culture, but not in relatively large-scale bioreactors.
Our device might aid in the scale-up of small-scale culture to bioreactors, as well as in the screening of culture-dependence of microorganisms, identification of useful metabolites, and screening for new microbes.
A small-scale culture was prepared by inoculating a single colony of S. cerevisiae into a medium containing 20 g L−1 glucose, 20 g L−1 peptone, and 10 g L−1 yeast extract.
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