宣泽生物在SCI期刊《Nanomedicine》发文——纳米孔检测技术推动医学进步

2017-09-11
阅读(


 

原文|《纳米医学》(Nanomedicine (Lond). 2017 Aug;12(16):1907-1909)

作者Farzin Haque,Shaoying Wang,Taoxiang Wu & Peixuan Guo

翻译|华昀
 

近年来,国际纳米科学和纳米孔道技术高速发展,并广泛应用于疾病检测及治疗,为建立有效的癌症诊断和治疗技术提供了新的契机。在此背景下,由深圳医药行业协会、深圳市龙华区经济促进局、深圳市龙华区科技创新局主办,深圳宣泽生物医药有限公司承办的2017首届国际纳米孔技术检测应用学术研讨会于三月底在深圳顺利召开,会议内容和成果得到了国内外的广泛关注和传播。继国内多家知名媒体跟踪报道后,国际著名学术期刊--纳米医学《Nanomedicine》(SCI影响因子4.7),在其最新一期期刊中(ISSN 1743-5889号),以“纳米孔检测技术推动医学进步”为标题,从学术角度报道了本次会议的内容和成果。现将报道的主要内容摘要翻译如下:

 

此次会议研讨了三大主题:1、利用纳米孔技术的独特性,推动医学进步;2、鼓励跨学科合作研究; 3、未来发展需要面对的挑战性。会议由The Ohio State University (俄亥俄州立大学)的RNA纳米生物和纳米医学中心主任及首席讲座教授郭培宣和中国科学院生物物理研究所著名教授张先恩共同主持。约有300多名来自各个国家、不同学科之间的学术研究人员、医院专家、政府领导和科学家参加了本次会议。中国“十三五”规划提出要大力发展基因组测序技术、高灵敏度检测技术、肿瘤靶向诊断和治疗方法。第十一届全国政协副主席白立忱、国家发改委宏观经济研究院副院长陈东琪、科技部发展战略研究院副院长王宏广,以及深圳市政府、深圳市科技创新委员会、卫生计划生育委员会、发展和改革委员会、龙华区政府的相关领导,共同出席了会议。

 

纳米孔是在基质上形成的。现有两种类型的纳米孔:由蛋白质或核酸组成的嵌入脂质或聚合物膜的生物孔,或者是在固体基质中的合成孔。检测方法是,当单个分子通过或与孔结合/相互作用的时候,电子监测通过孔的离子流。纳米孔平台在过去十年中迅速发展,在DNA和RNA测序、早期疾病诊断和各种其他应用方面取得了重大进展。大会报告集中在以下几个领域:中国政府政策支持、纳米孔测序、纳米孔道技术进展和早期疾病诊断。

 

来自华盛顿大学的Jen Gundlach教授介绍了使用MspA纳米孔道的DNA链测序;俄亥俄州立大学的Cythia Carnes副院长提到了快速低成本整体外泌体技术;宾夕法尼亚大学的Marija教授讨论了超薄基底、仪器设计和新电子,是怎样通过制造纳米带和场效应的提高纳米孔的时间和空间分辨率。清华大学药学院的白净卫教授展示了用于制造可扩展固态纳米孔器件和隧道间隙器件的侧壁转移工艺。特别是郭培宣教授介绍了使用从噬菌体phi29 DNA包装马达衍生而来的强大的纳米通道的研究进展。随后,来自深圳宣泽生物医药有限公司的王少英博士展示了纳米孔如何在单分子水平上实时检测肽氧化态。他的另一位同事FarzinHaque介绍了使用纳米孔捕获和指纹分析单分子的概念。phi29纳米孔内部通过突变修饰,用于捕获具有高特异性和灵敏度的单一化学物质。通过在phi29纳米孔道的末端设计多肽探针,可以在患者血清中存在许多污染物的情况下捕获特异性抗体。

 

多个学者和科学家在会上展示他们在该领域代表世界最尖端科学技术的最新研究成果。会议着重介绍了纳米孔的进步如何将基础科学转化为实用和临床应用的新途径。邀请的发言者举行了闭门圆桌会议,集思广益,共同努力解决这些课题。

 

参考文献:

 

Haque F, Wang S1, Wu T, Guo P. Advances in nanopore sensing promises to transform healthcare. Nanomedicine (Lond).  2017 Aug;12(16):1907-1909 

Nanopores are nanometer-sized holes in a substrate. There are two types of nanopores: biological pores composed of proteins or nucleic acids embedded in lipid or polymer membranes, and synthetic pores fabricated in solid substrates. The detection method is electrical monitoring of ion-flow through the pore, as single molecules pass or bind/interact with the pore. Both nanopore platforms have progressed rapidly over the last decade and significant advances have been made toward DNA and RNA sequencing, early disease diagnosis and a variety of other applications, as reported in a number of talks by leading researchers in the field. Oral presentations were organized into several themes: China government initiative, nanopore sequencing, technological advances, biophysical studies of macromolecule transport across nanopores and early disease diagnosis.

 

China government initiative

 

China's 13th Five-Year Plan has proposed to vigorously develop genome sequencing technologies for various diseases, highly-sensitive detection technologies, tumor targeted diagnosis and treatment methods. In response to this federal government initiative, local Shenzhen government sent several high-profile government leaders from Shenzhen municipality and Longhua District Government including Shenzhen Science and Technology Innovation Committee, Health and Family Planning Commission, and Development and Reform Commission. The leaders emphasized the need to continuously innovate to stay at the forefront of technology development and reinforced government resource support of enterprises working with cutting-edge technologies.

 

Nanopore sequencing

 

One of the major advantages of nanopore sequencing is the feasibility of sequencing the genome in a short time frame and under US$1000 per genome. The concept is rather simple: as a DNA strand traverses through the pore under an applied voltage, individual bases are identified based on modulations of ion current. However, there were several bottlenecks, such as speed of translocation, spatial and temporal resolution, biophysical properties of the pore and instrumentation. Jens Gundlach (University of Washington, USA) is leading the way of nanopore-based strand-sequencing using MspA nanopores. An enzyme (Phi29 DNA polymerase of helicase) is bound tightly to the single-stranded DNA and in the process, ratchets the DNA through the pore, one nucleobase at a time. However, base calling accuracy needs improvements as several bases in the narrow sensing region of the pore influence the electrical signal and also the enzyme turnover is stochastic. Gundlach's group is developing sophisticated bioinformatics approaches to address these issues toward more accurate nanopore sequencing. One prospective application is whole exosome sequencing as point of care technology. Cynthia Carnes (The Ohio State University, USA) highlighted the need for a rapid low-cost whole exosome sequencing technique that can be integrated with established clinical platforms to understand human arrhythmia pathophysiology and disease treatment.

 

Technological advances

 

Marija Drndic (University of Pennsylvania, USA) discussed how advances in ultra-thin solid substrates, device designs and new electronics have improved the temporal and spatial resolution of nanopores exemplified by fabrication of nanoribbons and field-effect transistors. Jingwei Bai (Tsinghua University, China) showed some recent developments in side-wall image-transfer process for fabricating scalable solid state nanopore devices and tunneling gap devices. Yitao Long (East China University of Science and Technology, China) combined optical and electrical readout signal using novel plasmonic nanopores. Enhanced scattering of light can be monitored through a nanopore on gold film using dark-field microscopy to characterize single molecules at unprecedented details. Shuo Huang (Nanjing University) demonstrated optical sensing of nucleic acids in a high-throughput manner. An array of 2500 single droplet interfaced bilayer can be generated in a micropatterned hydrogel chip for characterizing nucleic acid sequence specific binding events in parallel. Deqiang Wang (Chinese Academy of Sciences) developed a scalable method of fabricating graphene nanopores with a helium ion microscope. Compared with conventional nanofabrication techniques, this method is faster and allows precise control of size and shape of the synthetic pores.

 

Biophysical studies of macromolecule transport across nanopores

 

Meni Wanunu (Northeastern University, USA) highlighted fundamental properties of nanopores, such as pore charge, dimension, pressure and electric fields, and how these factors influence transport of charged polymers in small confinements. Peixuan Guo presented highlights of his research using a robust nanochannel derived from bacteriophage phi29 DNA packaging motor. The channel represents one of the largest biological pores embedded in a lipid membrane. The channel can translocate single stranded and double stranded nucleic acids as well as peptides of various composition and length. Studying heterogenous and transient systems is difficult using conventional biochemical techniques. Shaoying Wang (P&Z Biological Technology, NJ, USA) demonstrated how nanopores can detect peptide oxidation states in real-time at single molecule level. Similarly, Qing Zhao (Peking University, USA) used synthetic nanopores for determining the time-dependent kinetics of α-synuclein oligomerization that has been implicated in the pathogenesis of Parkinson's disease. Zuhong Lu (Southeast University) presented advances in developing a hybrid device composed of a solid state pore with a fixed protein machine to study the dynamic properties of a single protein using horseradish peroxidase as an example. Yitao Long (East China University of Science and Technology, China) showed that aerolysin nanopore can resolve 2-nt oligonucleotides and can monitor the cleavage of oligonucleotides by exonuclease-I. His lab also developed a simultaneous optical and electrical detection method for studying different conformations of fluorescently labeled DNA through quartz nanopores. Haichen Wu (Chinese Academy of Sciences) used DNA probes, such as aptamers for sensing metal ions, small organic molecules and biopolymers using α-hemolysin channels. Jia Geng (Sichuan University, China) developed synthetic analogs of biological membrane channels using carbon nanotubes and inserted them into a lipid bilayer as well as cell membrane to serve as a nanopore for transport of ions and biomolecules.

 

Early disease diagnosis

 

Early detection is critical for treating many types of diseases, including cancer, bacterial and viral infections, and cardiovascular diseases among others. Liquid biopsy of patient body fluids is a rapidly emerging method for detecting biomarkers. While sequencing of DNA is one approach relying on the target molecule passing through the pore, another approach is to functionalize the terminal ends of the pores with programmable probes that can bind a target molecule with high specificity and sensitivity. Farzin Haque (P&Z Biological Technology) presented the concept of capture and fingerprinting of single molecules using nanopores. Programmable receptors can be conjugated in the interior of phi29 nanopores for capturing single chemicals with high specificity and sensitivity. Peptide probes can also be engineered at the terminal ends of the phi29 channel to capture specific antibodies in the presence of many contaminants in the patient serum. Xinghua Lu (Chinese Academy of Sciences) demonstrated how synthetic nanopores can be used to detect DNA origami probes, bound to specific segments of DNA, for genomic diagnosis.

 

Conclusion

 

The conference highlighted how advances in nanopore are making new avenues for translating basic science into practical and clinical applications. A round table closed discussion session was held among the invited speakers to brainstorm fundamental challenges and discuss collaborative efforts to solve these challenges.

 

Financial & competing interests disclosure

 

F Haque and S Wang are employees of P&Z Biological Technology. P Guo is a consultant for Oxford Nanopore, Nanobiodelivery Pharmaceutical Co. Ltd and P&Z Biological Technology. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

 

No writing assistance was utilized in the production of this manuscript.
 

公司概况

宣泽生物成立于2016年6月,为全球第一家从事基于蛋白纳米孔道的单分子检测技术进行癌症超早期检测的公司。公司核心技术系基于郭培宣教授发明及持有的原创专利,并拥有其基于蛋白纳米孔道的单分子检测技术的全球独占许可。公司将持续加大科研投入,前期将陆续开发出灵敏度到单分子检测水平的多种癌症早期检测仪器及生物试剂,并与英国著名的纳米孔技术生物仪器公司保持紧密合作。

 

 

宣泽生物总部位于中国深圳市龙华区,在美国新泽西理工大学设立了全资子公司——P&Z BIOLOGICAL TECHNOLOGY LLC,与美国俄亥俄州大学正式签订了合同进行深度合作,依托世界级科研团队的加盟与智慧,开展核心技术的研发。公司在美国、中国均建有实验室,现已启动医疗器械市场准入前的各项准备工作,产品预计在2018年底面市,将前沿科学技术造福世人!
 


▲ 宣泽生物与OSU签署产品研发合作