Surface Patterning of Biomaterials Using the Tetrazine Ligation

在这项由大分子、超分子和纳米化学计划资助的奖项中，来自特拉华大学的Thomas Beebe博士、Joseph Fox博士和俄勒冈州立大学的Ryan Mehl博士将开发一种新的方法来创建生物分子表面的空间图案，包括梯度。该方法利用了福克斯博士开发的一种极其快速的生物正交反应--四嗪结扎法。拟议程序的基本新颖组成部分是能够控制附着在表面上的蛋白质的呈现。这将使用Mehl博士开发的一种方法来实现，在这种方法中，含有四嗪的非天然氨基酸被特定地展示在蛋白质表面暴露的位置。将这两种元素结合在一起，可以使用微接触打印技术在表面打印可点击的反式环辛烯，然后与四嗪修饰的蛋白质和蛋白质结构域发生反应，从而产生微米到厘米的图案和梯度。这项提议的三个主要目标是：(A)从反式环辛烯开始，合成和表征表面固定蛋白质的梯度和模式；(B)开发表面光化学图案化的方法；(C)用于定向纤维连接蛋白结构域的神经元引导的许可底物。该奖项的所有目标都利用了受控蛋白质呈现这一新颖元素。由于许多新兴生物技术依赖于生物分子在表面的附着，最好是处于生物活性状态，该项目可以极大地提高这些新兴技术的检测极限、读出速度、保质期和一般功能。该奖项包括一个教育和推广计划，涉及高中生、他们的老师、本科生、研究生和博士后研究人员，他们都在特拉华大学和俄勒冈州立大学的三个不同的研究小组之间合作，使用有机合成、表面化学和分析以及生物有机化学。该奖项还将支持对当地非裔美国高中教师威尔明顿友谊学校的Loraine Snead女士的外展努力。斯奈德开发了一门高中课程，向学生传授现代分析和光谱技术。在过去的三年里，福克斯博士一直与斯奈德合作，支持她的课程创新。

In this award funded by the Macromolecular, Supramolecular, and Nanochemistry Program, Drs. Thomas Beebe, Joseph Fox (both from the University of Delaware) and Ryan Mehl of Oregon State University will develop a novel method for the creation of spatial patterns, including gradients, of biological molecules on surfaces. The approach utilizes the tetrazine ligation, an extremely rapid bioorthogonal reaction developed by Dr. Fox. The essential novel component of the proposed program is the ability to control the presentation of proteins that are attached to a surface. This will be achieved using a method developed by Dr. Mehl, in which tetrazine-containing unnatural amino acids are site-specifically displayed at surface-exposed positions in proteins. Putting these two elements together results in the ability to create micrometer to centimeter patterns and gradients using microcontact printing of the clickable trans-cyclooctene on the surface, followed by reaction with tetrazine-modified proteins and protein domains. The three main objectives of this proposal are: (a) synthesis and characterization of gradients and patterns of immobilized proteins on surfaces, starting from trans-cyclooctene; (b) development of methods for photochemically patterning surfaces; (c) permissive substrates for neuron guidance with oriented fibronectin domains. All of the award's objectives make use of the novel element of controlled protein presentation. Since many emerging biotechnologies rely on the attachment of biomolecules to surfaces, preferably in a bioactive state, this project can greatly enhance detection limits, readout speed, shelf-life, and general functionality of these emerging technologies.This award features an education and outreach plan involving high school students, their teachers, undergraduate students, graduate students and postdoctoral researchers, all working together between three different research groups using organic synthesis, surface chemistry and analysis, and bioorganic chemistry at the University of Delaware and at Oregon State University. This award will also support outreach efforts to a local African-American high school teacher, Ms. Loraine Snead of Wilmington Friends School. Ms. Snead has developed a high school curriculum that teaches students modern analytical and spectroscopic techniques. Over the past 3 years, Dr. Fox has worked with Ms. Snead to support her curricular innovations.