High-throughput microfluidics for the screening of yeast libraries
Artikel i vetenskaplig tidskrift, 2018

Cell factory development is critically important for efficient biological production of chemicals, biofuels, and pharmaceuticals. Many rounds of the Design–Build–Test–Learn cycles may be required before an engineered strain meeting specific metrics required for industrial application. The bioindustry prefer products in secreted form (secreted products or extracellular metabolites) as it can lower the cost of downstream processing, reduce metabolic burden to cell hosts, and allow necessary modification on the final products, such as biopharmaceuticals. Yet, products in secreted form result in the disconnection of phenotype from genotype, which may have limited throughput in the Test step for identification of desired variants from large libraries of mutant strains. In droplet microfluidic screening, single cells are encapsulated in individual droplet and enable high-throughput processing and sorting of single cells or clones. Encapsulation in droplets allows this technology to overcome the throughput limitations present in traditional methods for screening by extracellular phenotypes. In this chapter, we describe a protocol/guideline for high-throughput droplet microfluidics screening of yeast libraries for higher protein secretion. This protocol can be adapted to screening by a range of other extracellular products from yeast or other hosts.

Droplet microfluidics

High-throughput screening

Systems biology

Yeast cell factories

Protein secretion

Random mutagenesis

Författare

Mingtao Huang

Chalmers, Biologi och bioteknik, Systembiologi

H. N. Joensson

The Royal Institute of Technology (KTH)

Jens B Nielsen

Chalmers, Biologi och bioteknik, Systembiologi

Methods in molecular biology (Clifton, N.J.)

1064-3745 (ISSN)

Vol. 1671 307-317

Ämneskategorier

Biokemi och molekylärbiologi

DOI

10.1007/978-1-4939-7295-1_19