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METHODS OF LABELING TRANSIENTLY EXPRESSED PROTEINS IN LARGE-SCALE EUKARYOTIC CELL CULTURES

机译:在大型真核细胞培养物中标记瞬时表达蛋白的方法

摘要

The present invention is based on observations that transiently transfecting large- scale eukaryotic cell cultures with a polynucleotide encoding a protein of interest can be used to rapidly produce the large quantities of labeled proteins required for various biochemical techniques such as spectroscopy, microscopy, and crystallography, and applications including protein structure determination, protein tracing and/or localization, diagnostic and therapeutic applications, and affinity experiments. Thus, the present invention provides methods for rapidly producing large quantities of labeled proteins by using transient transfection of large-scale eukaryotic cell cultures, which are then grown in a chemically defined labeling medium that includes labeled amino acids. The present invention is also directed to methods of using the labeled proteins produced by the novel labeling methods for use in various techniques.
机译:本发明基于以下观察结果:用编码目的蛋白的多核苷酸瞬时转染大规模真核细胞培养物可用于快速生产各种生化技术(例如光谱学,显微镜和晶体学)所需的大量标记蛋白,应用包括蛋白质结构测定,蛋白质追踪和/或定位,诊断和治疗应用以及亲和力实验。因此,本发明提供了通过使用大规模真核细胞培养物的瞬时转染来快速产生大量标记蛋白的方法,然后将其在包含标记氨基酸的化学确定的标记培养基中生长。本发明还涉及使用由新颖的标记方法产生的标记蛋白质的方法,以用于各种技术。

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