Surface Patterning of Biomaterials Using the Tetrazine Ligation

在这个由大分子、超分子和纳米化学项目资助的奖项中，托马斯毕比博士、约瑟夫福克斯博士（均来自特拉华州大学）和俄勒冈州州立大学的瑞安梅尔博士将开发一种新的方法，用于在表面上创建生物分子的空间图案，包括梯度。该方法利用四嗪连接，这是Fox博士开发的一种非常快速的生物正交反应。 所提出的程序的基本新组成部分是控制附着在表面上的蛋白质的呈现的能力。 这将使用Mehl博士开发的方法来实现，其中含有四嗪的非天然氨基酸在蛋白质的表面暴露位置处位点特异性地展示。将这两种元素放在一起，能够使用表面上可点击的反式环辛烯的微接触印刷产生微米至厘米的图案和梯度，然后与四嗪修饰的蛋白质和蛋白质结构域反应。该提案的三个主要目标是：（a）从反式环辛烯开始，合成和表征表面上固定化蛋白质的梯度和图案;（B）开发用于光化学图案化表面的方法;（c）具有定向纤连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的新颖元素。由于许多新兴的生物技术依赖于生物分子附着在表面上，最好是在生物活性状态下，这个项目可以大大提高检测限，读出速度，保质期，和这些新兴技术的一般功能。这个奖项的特点是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，他们在特拉华州大学和俄勒冈州州立大学的三个不同的研究小组之间进行合作，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洲裔美国高中教师、威尔明顿朋友学校的洛琳·斯内德女士的外联工作。Snead女士开发了高中课程，教授学生现代分析和光谱技术。 在过去的3年里，福克斯博士与斯尼德女士合作，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.