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Microbead display by in vitro compartmentalisation: selection for binding using flow cytometry
Author(s) -
Sepp Armin,
Tawfik Dan S,
Griffiths Andrew D
Publication year - 2002
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(02)03740-7
Subject(s) - microbead (research) , flow cytometry , microbiology and biotechnology , in vitro , chemistry , gene , dna , biology , computational biology , biochemistry
In vitro compartmentalisation in an emulsion was used to physically link proteins to the DNA that encodes them via microbeads. These microbeads can be selected for catalysis, or, as demonstrated here, for binding. Genes encoding a peptide containing an epitope (haemagglutinin) were enriched to near purity from a 10 6 ‐fold excess of genes encoding a different peptide by two rounds of selection using flow cytometry, indicating ∼1000‐fold enrichment per round. Single beads can be isolated using flow sorting and the single gene on the bead amplified by polymerase chain reaction. Hence, the entire process can be performed completely in vitro.