New detection modes for droplet microfluidics

液滴微流控的新检测模式

• 批准号: BB/K013629/1
• 负责人: Florian Hollfelder
• 金额: $ 11.19万
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2013
• 资助国家: 英国
• 起止时间: 2013 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FK013629%2F1
• 关键词: New; detection; modes; droplet; microfluidics

This project is based on the idea that the classical test tube in experimental biology can be replaced by miniature water-in-oil droplets. The droplet is the equivalent of an ultra-small test tube. Droplet volumes are typically in the femto- to nanolitre range. Furthermore, up to 10e10 droplet reactors fit into a millilitre-sized tube. Importantly, an equivalent number of experiments can be carried out simultaneously. The key idea is that the droplet compartment combines the functional molecule with information on its identity and a readout of its function. Thus the droplet contains everything needed to assess and decode a particular experiment or profile a library member. This system is ideally suited for directed evolution, because genotype and phenotype are combined in the droplet. We have recently performed directed protein evolution in this format (Kintses, B. et al & Hollfelder, F. Chemistry & Biology 2012, 19(8):1001-9) and now propose to add another detection mode for assays that are based on generation of a chromophore, to allow a wider range of biological functions to be screened.

这个项目是基于这样的想法：实验生物学中的经典试管可以被微型油包水滴所取代。液滴相当于一个超小型试管。液滴体积通常在飞升到纳升范围内。此外，多达10e10个液滴反应器可装入一毫升大小的管中。重要的是，可以同时进行相同数量的实验。关键的想法是，液滴隔间将功能分子与其身份信息和功能读数结合起来。因此，液滴包含了评估和解码特定实验或描述库成员所需的一切。该系统非常适合定向进化，因为基因型和表型在液滴中结合。我们最近以这种形式进行了定向蛋白质进化（Kintses， B. et al . & Hollfelder， F. Chemistry & Biology 2012, 19(8):1001-9），现在提议增加另一种检测模式，用于基于发色团生成的分析，以允许更广泛的生物功能进行筛选。

项目成果

A method to quantify FRET stoichiometry with phasor plot analysis and acceptor lifetime ingrowth.

• DOI: 10.1016/j.bpj.2015.01.012
• 发表时间: 2015-03-10
• 期刊: BIOPHYSICAL JOURNAL
• 影响因子: 3.4
• 作者: Chen, WeiYue;Avezov, Edward;Schlachter, Simon C.;Gielen, Fabrice;Laine, Romain F.;Harding, Heather P.;Hollfelder, Florian;Ron, David;Kaminski, Clemens F.
• 通讯作者: Kaminski, Clemens F.

Improved RAD51 binders through motif shuffling based on the modularity of BRC repeats

通过基于 BRC 重复模块性的基序改组改进 RAD51 结合物

• DOI: 10.1101/2020.05.14.097071
• 发表时间: 2020
• 作者: Lindenburg L

Quantitative Affinity Determination by Fluorescence Anisotropy Measurements of Individual Nanoliter Droplets.

• DOI: 10.1021/acs.analchem.6b02528
• 发表时间: 2017-01-17
• 期刊: Analytical chemistry
• 影响因子: 7.4
• 作者: Gielen F;Butz M;Rees EJ;Erdelyi M;Moschetti T;Hyvönen M;Edel JB;Kaminski CF;Hollfelder F
• 通讯作者: Hollfelder F

Improved RAD51 binders through motif shuffling based on the modularity of BRC repeats.

通过基于 BRC 重复模块性的基序改组改进了 RAD51 结合物。

• DOI: 10.17863/cam.74358
• 发表时间: 2021
• 作者: Lindenburg L

Ultrahigh-throughput-directed enzyme evolution by absorbance-activated droplet sorting (AADS)

通过吸光度激活液滴分选 (AADS) 进行超高通量定向酶进化

• DOI: 10.17863/cam.6859
• 发表时间: 2016
• 作者: Gielen F