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Titre reached up to 3.05 μg mL−1 in small scale expression.
Based on the analysis of cell lysate from the small scale expression of five antibodies, we have determined that best expression levels can be obtained using only two sets of conditions: for PA38, PA64 and 4G2, optimal expression was observed from the high copy plasmid, induced at OD600 1.0 with 20 ng/ml inducer.
After yeast transformation, a small scale expression assay was performed.
A random selection of resulting colonies was screened for small scale expression.
Western blots from the small scale expression show a clear difference in expression levels among the different clones.
Prior to use for requested target proteins, every new vector was validated with eGFP for cloning and small scale expression in the respective host.
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Small-scale expression conditions were tested for all these constructs.
The truncated TRAM constructs were designed based on secondary structure predictions and screened by small-scale expression.
We emphasize technologies that have been evaluated and implemented in our laboratory, including innovative molecular biology and expression vectors, small-scale expression screening strategies and the automation of parallel and multidimensional chromatography.
Small-scale expression experiments were carried out to select the optimal conditions for solubility of the expressed proteins.
Small-scale expression screening with soluble fusion tags identified many conditions that yielded soluble expression.
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