Directed Evolution of Formylglycine-generating Enzyme to Build an Optimal Platfor

描述（由申请人提供）：甲酰基甘氨酸生成酶（FGE）识别并特异性作用于五肽（CXPXR），将半胱氨酸氧化为不寻常的含醛甲酰基甘氨酸残留物（fGly）。将该序列插入重组蛋白中，就有可能转化为fGly。小的FGE靶序列，被称为“醛标签”，是红木生物科学技术的基础，可以产生同质的、增强的生物治疗药物。该平台提供了在几乎任何蛋白质上产生有用的翻译后修饰所需的位点特异性和化学灵活性。这种将新的化学功能附加到感兴趣的生物实体上的能力是开发和优化新疗法的一个非常强大的平台。随着该技术的应用探索，在FGE识别的共识序列中具有灵活性将是有利的，以便允许对尽可能多的蛋白质靶标进行修饰，理想情况下是在其内源序列中已经存在的位点上。一组FGE酶/标签对将是一个令人难以置信的强大平台，并产生有价值的知识产权。新的标签序列将被设计为最小化对蛋白质序列的扰动，最大化FGE特异性活性，从而实现优化的偶联。这种新的标签技术将产生具有最小抗原性和增强药代动力学的生物偶联物。为此，红木生物科学公司将开展一项研究计划，利用酵母细胞表面蛋白展示的定向进化方法，产生识别独特和新颖底物的FGE变体。

DESCRIPTION (provided by applicant): The formylglycine generating-enzyme (FGE) recognizes and acts specifically on a pentapeptide (CXPXR), oxidizing the cysteine to an unusual aldehyde bearing formylglycine residue (fGly). This conversion to fGly is possible when the sequence is inserted into recombinant proteins. The small FGE target sequence, termed the "aldehyde tag", anchors the Redwood Bioscience technology that can generate homogenous, enhanced biotherapeutics. This platform provides the site specificity and chemical flexibility needed to generate useful post translational modifications on virtually any protein. This ability o affix novel chemical functionalities to a biological entity of interest is an immensely powerful platform for the development and optimization of new therapeutics. As applications of this technology are explored it will be advantageous to have flexibility in the consensus sequence recognized by FGE, so as to permit the modification of as many protein targets as possible, ideally on sites that exist already in their endogenous sequences. A panel of FGE enzyme/tag pairs would be an incredibly powerful platform and generate valuable intellectual property. The new tag sequences will be designed to minimize perturbation of the protein sequence and maximize FGE specific activity, thereby enabling optimized conjugation. This new tag technology will generate bioconjugates with minimal antigenicity and enhanced pharmacokinetics. Towards this end, Redwood Bioscience will embark on a research program using a directed evolution approach with yeast cell surface protein display to generate FGE variants that recognize unique and novel substrates.