We describe a mammalian expression construct (SPLICELECT™) that allows the redirection of a proportion of a secreted protein onto the cell surface using alternative splicing: whereas the majority of the RNA is spliced into a transcript encoding a secreted protein, a weak splice donor site yields a secondary transcript encoding, in addition, a C-terminal transmembrane domain. The different sequence elements can be modified in order to modulate the level of cell surface display and of secretion in an independent manner. In this work, we demonstrated that the cell surface display of stable cell lines is correlated with the level of the secreted protein of interest, but also with the level of heterodimerization in the case of a bispecific antibody. It was also shown that this construct may be useful for rapid screening of multiple antibody candidates in binding assays following transient transfection. Thus, the correlation of product quantity and quality of the secreted and of membrane-displayed product in combination with the flexibility of the construct with regards to cell surface display/secretion levels make SPLICELECT™ a valuable tool with many potential applications, not limited to industrial cell line development or antibody engineering.

中文翻译：

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SPLICELECT™：一种基于可变剪接的适应性细胞表面展示技术，可在单细胞水平上对分泌产物进行定性和定量预测。

我们描述了一种哺乳动物表达构建体（SPLICELECT™），它允许使用替代性剪接将一部分分泌的蛋白质重定向到细胞表面：而大部分RNA剪接成编码分泌蛋白的转录本，剪接供体位点弱产生编码C末端跨膜结构域的二级转录物。可以修饰不同的序列元件，以便以独立的方式调节细胞表面展示和分泌的水平。在这项工作中，我们证明了稳定细胞系的细胞表面展示与所关注蛋白质的分泌水平相关，但在双特异性抗体的情况下，也与异二聚化水平相关。还显示出该构建体可用于瞬时转染后的结合测定中快速筛选多种抗体候选物。因此，分泌物和膜展示产品的产品数量和质量的相关性，以及构建体在细胞表面展示/分泌水平方面的灵活性，使SPLICELECT™成为具有许多潜在应用的有价值的工具，不仅限于工业应用细胞系开发或抗体工程。