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计划的支持下，加州大学欧文分校的科恩教授正在开发和应用新的光学技术，这些技术可以检测、表征和实时监控单个聚合物纳米颗粒的尺寸和组成。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，从根本上说是重要和有趣的。康恩教授开发了更好的化学方法来提高这些纳米颗粒表征的一致性、可靠性和稳定性。科恩教授的研究成果可能会推动传感器开发、药物输送和疾病诊断领域的技术进步。科恩教授的项目为参与的学生提供了一系列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，这笔赠款还支持教育和推广项目，以吸引资金不足学校的高中生和对科学感兴趣的中学生进入STEM领域。单纳米颗粒近红外表面等离子共振成像(SPRI)显微镜是一种基于折射率的新方法，用于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力吸收和释放。SPRI是一种成熟的基于折射率的技术，用于监测分子和纳米颗粒在金薄膜上的吸附。这里将SPRI显微镜的应用扩展到单纳米颗粒测量，通过测量由于组成变化而发生的单纳米颗粒折射率的变化，可以实时监测生物分子(包括单纳米颗粒平均和分布测量)的吸收和释放。合成和研究了多种改性N-异丙基丙烯酰胺(NIPAM)水凝胶纳米粒，它们可以吸收和释放不同的蛋白质、凝集素、核酸、多肽和药物。三个具体的研究工作包括：(I)NIPAM水凝胶纳米颗粒的合成和表征，其结合点击化学位点以连接糖、肽或DNA以用于治疗药物、蛋白质和RNA的生物亲和力吸收和释放；(Ii)DNA-交联型NIPAM(N-异丙基丙烯酰胺)水凝胶纳米颗粒的合成和表征，其结合DNA适配子和DNAzyme；以及(Iii)单纳米颗粒测量NIPAM水凝胶纳米颗粒的pH和温度引起的释放各种货物(例如，肽、酶、DNA、这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势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.