微液滴高通量筛选方法的研究与应用进展

秦伟彤; 杨广宇

doi:10.12211/2096-8280.2023-033

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微液滴高通量筛选方法的研究与应用进展

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- 微液滴高通量筛选方法的研究与应用进展
- Research and application progress of microdroplets high throughput screening methods
- 合成生物学 2023年4卷第5期页码：966-979
- 作者机构：
  
  上海交通大学生命科学技术学院，微生物代谢国家重点实验室，上海 200240
- 作者简介：
  
  [ "秦伟彤（1992—），女，博士。研究方向为酶的定向进化、微液滴超高通量筛选方法的建立。E-mail：qinweitong122@163.com" ]
  [ "杨广宇（1980—），男，研究员，博士生导师。研究方向为酶分子改造、超高通量筛选方法的建立；酶分子机制解析；体外合成生物学研究。E-mail：yanggy@sjtu.edu.cn" ]
- 基金信息：
  
  国家重点研发计划(2018YFE0200501;2020YFA0907900);国家自然科学基金(32030063);广东省重点领域研发计划(2022B1111050001);天津市合成生物技术创新能力提升行动;上海交通大学特区计划(21TQ1400210)
- DOI：10.12211/2096-8280.2023-033
  中图分类号： Q818
- 收稿：2023-04-24，
  
  修回：2023-06-20，
  
  纸质出版：2023-10-31
- 稿件说明：
移动端阅览
秦伟彤，杨广宇. 微液滴高通量筛选方法的研究与应用进展［J］. 合成生物学， 2023， 4（5）： 966-979

QIN Weitong， YANG Guangyu. Research and application progress of microdroplets high throughput screening methods［J］. Synthetic Biology Journal， 2023， 4（5）： 966-979
秦伟彤，杨广宇. 微液滴高通量筛选方法的研究与应用进展［J］. 合成生物学， 2023， 4（5）： 966-979 DOI： 10.12211/2096-8280.2023-033.

QIN Weitong， YANG Guangyu. Research and application progress of microdroplets high throughput screening methods［J］. Synthetic Biology Journal， 2023， 4（5）： 966-979 DOI： 10.12211/2096-8280.2023-033.

摘要

在单细胞层面对生物功能进行高通量的分析和分选是对关键基因、元件、途径与细胞工厂进行优化的重要技术。基于微液滴的筛选方法因其低成本、超高通量等优势，已被广泛应用于生物、医药、食品和工业等各个领域。本文针对目前主流的荧光激活的液滴分选、吸光度激活的液滴分选，以及无标记液滴分选等微液滴筛选设备的进展进行综述，主要包括基于质谱、拉曼、核磁共振、电化学、图像识别等。并总结了近年来微液滴筛选设备在酶进化、微生物育种等领域应用成功的案例。此外还对不同的微液滴筛选设备的优势与面临的挑战进行了讨论，未来各种新的荧光探针的开发以及质谱等非标记检测方法的进一步发展，将是微液滴筛选设备的主要发展方向，在蛋白质工程、抗体工程、细胞分选及临床研究等方面具有重要的应用潜力。

Abstract

High throughput analysis and sorting of biological functions at the single-cell level is an important technology for optimizing the performance key genes

elements

pathways

and cell factories. The screening method based on microdroplet has been widely applied in various fields such as biology

medicine

food

and industry due to its advantages of low cost and ultra-high throughput. Traditional droplet sorting includes droplet generation

incubation

operation

and sorting. For the first three steps

there have been many technological advances in the last decade. The major limitation is in the sorting step

whose frequency and diversity restrict the sorting efficiency and target scope. According to the different principles of detection signals

droplet sorting technology is mainly divided into labeled and unlabeled sorting method. This article mainly reviews the progress of microdroplet screening equipment based on mainstream fluorescence-activated droplet sorting (FADS) to detect fluorescence signals

absorbance-activated droplet sorting (AADS) to detect UV/visible light absorption changes

and unlabeled droplet sorting

such as mass spectrometry

Raman spectrometry

nuclear magnetic resonance

electrochemistry

and image recognition. This article summarized the successful cases of microdroplet screening equipment applied in the fields of enzyme evolution and microbial breeding in the past 5 years. In addition

we also discussed the advantages and challenges faced by different microdroplet screening devices

and pointed out that the development of various new fluorescent probes and the further development of unlabeled detection methods such as mass spectrometry in the future will be the main development direction of microdroplet screening equipment. Although FADS remains the primary choice for cell sorting

the development of other microfluidic sorting devices has further expanded the application range of microfluidic sorting devices. Its high screening throughput and independent reaction environment provide a new technological platform for research in different fields

including protein engineering

antibody screening

sorting of different types of cells

and clinical directions.

关键词

Keywords

references

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