Flow-Based Single Cell Deposition for High-Throughput Screening of Protein Libraries

The identification and engineering of proteins having refined or novel characteristics is an important area of research in many scientific fields. Protein modelling has enabled the rational design of unique proteins, but high-throughput screening of large libraries is still required to identify prot...

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Bibliographic Details
Published in:PloS one 2015-11, Vol.10 (11), p.e0140730-e0140730
Main Authors: Stowe, Cassandra, Pizzey, Arnold, Kalber, Tammy, Badar, Adam, Lythgoe, Mark, Pule, Martin
Format: Article
Language:English
Subjects:
Bacteria
Bacteria - isolation & purification
Bacteria - metabolism
Cancer
Clinical trials
Colonies
Computational Biology
Deposition
Flow cytometry
Flow Cytometry - instrumentation
Flow Cytometry - methods
Fluorescence
Format
Green Fluorescent Proteins - genetics
Green Fluorescent Proteins - metabolism
Growth media
Hematology
High-throughput screening
High-Throughput Screening Assays
Hypotheses (Scientific method)
Interrogation
Medical research
Medicine
Microscopy
Photinus pyralis
Pichia - isolation & purification
Pichia - metabolism
Proteins
Proteins - genetics
Proteins - metabolism
Screening
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Summary:The identification and engineering of proteins having refined or novel characteristics is an important area of research in many scientific fields. Protein modelling has enabled the rational design of unique proteins, but high-throughput screening of large libraries is still required to identify proteins with potentially valuable properties. Here we report on the development and evaluation of a novel fluorescent activated cell sorting based screening platform. Single bacterial cells, expressing a protein library to be screened, are electronically sorted and deposited onto plates containing solid nutrient growth media in a dense matrix format of between 44 and 195 colonies/cm2. We show that this matrix format is readily applicable to machine interrogation (
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0140730