Growth of Hybrid Polymeric Nanostructures for Enzyme-Free Amplified Protein Imaging

蛋白质在人体中起着至关重要的作用。然而，由于它们通常在低浓度下被发现，它们可能很难检测到。克服这一挑战不会推进基础研究，也不会大大提高诊断质量和新疗法的发现。该项目的目标是开发一种新的生物传感纳米技术来放大目标蛋白质的信号。用这项新技术可以检测到用传统方法无法检测到的蛋白质。不同水平的学生将通过实践研究经验进行培训。通过宾夕法尼亚州立大学的各种外展活动，全国范围内代表性不足的学生将被招募到这一计划中。通过这个项目产生的新知识将被整合到为本科生和研究生提供的课程中。此外，这些知识将通过高知名度的期刊出版物和会议报告广泛传播。蛋白质检测在几乎所有与生物医学科学和临床诊断相关的领域都是至关重要的。然而，现有的蛋白质原位成像和传感方法往往存在信号强度弱、色素/纳米颗粒沉积和块状试剂扩散缓慢等问题。本项目的目标是探索一种新的方法，通过原位生长生物分子纳米结构来进行蛋白质成像中的无酶信号放大。探索这一新方法的成功将解决现有蛋白质检测方法面临的尚未满足的关键需求。在提出的新方法中，每个蛋白质靶标将由携带大量荧光团的超分子成像纳米结构表示。值得注意的是，这种纳米结构将被原位生长以用于成像，而不需要使用任何酶。因此，它不仅将提供信号放大，还将避免潜在的酶或纳米颗粒相关的问题。这项拟议的研究有三个任务：1)了解DNA分子的组成和结构如何决定DNA纳米结构的形成；2)合成和表征基于DNA的杂化超分子纳米结构；3)评估该方法在蛋白质成像中的有效性。这一项目的完成将产生广泛的影响。首先，它将推进蛋白质传感的方法学，使需要高灵敏度蛋白质成像的各种生物和生物医学研究领域受益。其次，对DNA聚合的基本了解将为开发各种基于DNA的生物传感器和材料提供知识基础。第三，由于临床活检的体积往往很小，这种无酶信号放大成像方法也具有提高临床诊断的巨大潜力。不同层次的人力资源开发将进一步加强广泛的影响，通过宾夕法尼亚州立大学的各种外展计划积极招募全国代表性不足的学生，在知名期刊上发表论文，并通过受邀的研讨会和会议演讲传播知识。该奖项反映了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。

Proteins play essential roles in the body. However, because they have typically are found in low concentration, they can be very challenging to detect. Overcoming this challenge will not advance basic research and greatly improve the quality of diagnosis and the discovery of new therapeutics. The objective of this project is to develop a novel biosensing nanotechnology to amplify the signals of target proteins. The proteins that cannot be detected using traditional methods would be detected with this new technology. Students at different levels will be trained through hands-on research experience. Nationwide underrepresented students through diverse outreach activities at Penn State will be recruited to this program. The new knowledge to be generated through this program will be integrated into the classes offered to undergraduate and graduate students. Moreover, the knowledge will be broadly disseminated through high-profile journal publications and conference presentations. Protein examination is pivotal to virtually all fields related to biomedical sciences and clinical diagnostics as well. However, existing methods for in situ protein imaging and sensing often suffer from problems such as weak signal intensity, pigment/nanoparticle deposition and slow diffusion of bulky agents. The objective of this project is to explore a novel method for enzyme-free signal amplification in protein imaging via in situ growth of biomolecular nanostructures. The success of exploring this new method will address the critical unmet needs faced by existing methods for protein detection. In the proposed new method, each protein target will be represented by a supramolecular imaging nanostructure that carries a large number of fluorophores. Notably, the nanostructure will be grown in situ for imaging without the need of using any enzyme. Thus, it will not only provide signal amplification but also avoid potential enzyme- or nanoparticle-associated problems. The proposed research is designed with three tasks: 1) to understand how the compositions and structures of DNA molecules determine the formation of DNA nanostructures; 2) to synthesize and characterize hybrid DNA-based supramolecular nanostructures; and 3) to evaluate the effectiveness of this method in protein imaging. The accomplishment of this project will make broad impacts. Firstly, it will advance the methodology of protein sensing, benefiting a variety of biological and biomedical research areas that require highly sensitive protein imaging. Secondly, the fundamental understanding of DNA polymerization will provide a knowledge basis for the development of various DNA-based biosensors and materials. Thirdly, as clinical biopsies often have very small volumes, this enzyme-free signal amplification imaging method also holds great potential of improving clinical diagnostics. The broad impacts will be further strengthened by the development of human resources at different levels with active recruitment of nationwide underrepresented students through a diverse array of Penn State outreach programs, the publication of papers in high-profile journals and the dissemination of knowledge through invited seminars and conference presentations.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.