Single Nanoparticle SPR Imaging Measurements of Bioaffinity Uptake and Release in Polymer and DNA-Cross-linked Nanoparticles

在化学系化学测量和成像项目的支持下，加州大学欧文分校的Corn教授正在开发和应用新的光学技术，这些技术可以真实的实时检测、表征和监测单个聚合物纳米颗粒的大小和组成。了解多孔聚合物纳米颗粒如何吸收，运输，然后释放生物活性分子，如核酸和蛋白质是非常重要和有趣的。Corn教授开发了更好的化学方法来提高这些纳米颗粒表征的均匀性，可靠性和稳定性。玉米教授的研究成果可能会导致传感器开发，药物输送和疾病诊断领域的技术进步。玉米教授的项目为参与的学生提供了不同学科的研究经验：聚合物材料和表征，纳米材料，激光显微镜，生物传感器和生物技术。培养具有如此广泛技能的年轻科学家对于工业界和学术界成功应对未来多学科技术挑战至关重要。除了研究之外，该补助金还支持吸引资金不足学校的高中生和对科学感兴趣的中学生到STEM领域的教育和推广计划。单纳米颗粒近红外表面等离子体共振成像（SPRI）显微镜是一种基于折射率的新方法，可以量化各种生物分子在单个多孔聚合物纳米颗粒中的生物亲和性吸收和释放。SPRI是一种已建立的基于折射率的技术，用于监测分子和纳米颗粒吸附到金薄膜上。 SPRI显微镜的应用在这里扩展到单个纳米颗粒测量，可以通过测量由于组成变化而发生的单个纳米颗粒折射率的变化来真实的监测生物分子的摄取和释放（单个纳米颗粒平均值和分布测量）。合成并研究了各种改性的N-异丙基丙烯酰胺（NIPAm）水凝胶纳米颗粒，其可以摄取和释放不同的蛋白质、凝集素、核酸、肽和药物。 三项具体的研究工作包括：（i）NIPAm水凝胶纳米颗粒的合成和表征，所述NIPAm水凝胶纳米颗粒并入点击化学位点以连接糖、肽或DNA，用于治疗剂、蛋白质和RNA的生物亲和性摄取和释放，掺入DNA适体和DNA酶的（N-异丙基丙烯酰胺）水凝胶纳米颗粒，和（iii）NIPAm水凝胶纳米颗粒中pH和温度诱导的形态变化的单纳米颗粒测量，所述NIPAm水凝胶纳米颗粒释放各种货物（例如，该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

With support from the Chemical Measurement and Imaging Program in the Division of Chemistry, Professor Corn at the University of California, Irvine, is developing and applying new optical techniques that can detect, characterize and monitor in real time the size and composition of single polymer nanoparticles. Understanding how porous polymer nanoparticles uptake, transport, and then release biologically active molecules such as nucleic acids and proteins are fundamentally important and interesting. Professor Corn develops better chemical methods to improve the uniformity, reliability, and stability of characterization of these nanoparticles. Professor Corn's research outcome could lead to technological advances in the fields of sensor development, drug delivery, and disease diagnostics. Professor Corn's project provides participating students with research experience in a diverse set of disciplines: polymer materials and characterization, nanomaterials, laser microscopy, biosensors and biotechnology. Development of young scientists with such a wide range of skills is essential for both industry and academia to successfully address future multi-disciplinary technological challenges. In addition to research, this grant supports education and outreach programs that attract high school students from underfunded schools and middle school female students interested in science to the STEM fields.Single nanoparticle near-infrared surface plasmon resonance imaging (SPRI) microscopy is developed as a novel refractive index-based method to quantify the bioaffinity uptake and release of various biomolecules into individual porous polymeric nanoparticles. SPRI is an established refractive index-based technique for monitoring molecular and nanoparticle adsorption onto gold thin films. The application of SPRI microscopy is expanded here to single nanoparticle measurements that can monitor the uptake and release of biomolecules (both single nanoparticle averages and distribution measurements) in real time by measuring changes in the single nanoparticle refractive index that occur due to changes in composition. A variety of modified N-isopropylacrylamide (NIPAm) hydrogel nanoparticles that can uptake and release different proteins, lectins, nucleic acids, peptides, and pharmaceuticals are synthesized and studied. Three specific research efforts include: (i) the synthesis and characterization of NIPAm hydrogel nanoparticles that incorporate click chemistry sites to attach sugars, peptides, or DNA for the bioaffinity uptake and release of therapeutics, proteins, and RNA, (ii) the synthesis and characterization of DNA-cross-linked NIPAm (N-isopropylacrylamide)hydrogel nanoparticles that incorporate DNA aptamers and DNAzymes, and (iii) single-nanoparticle measurements of pH and temperature-induced morphological changes in NIPAm hydrogel nanoparticles that release various cargo (e.g., peptides, enzymes, DNA, and gold nanoparticles).This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.