Continuous affinity-based selection: rapid screening and simultaneous amplification of bacterial surface-display libraries.
Biochem J
; 357(Pt 3): 779-85, 2001 Aug 01.
Article
em En
| MEDLINE
| ID: mdl-11463348
ABSTRACT
A new method for continuous biopanning has been developed. We have combined the power of affinity chromatography with the fecundity of bacteria in a unique process that mimics clonal selection. Mixed populations of bacteria were applied to a fermenter containing the immobilized ligand of interest. Bacteria retained in this affinity fermenter were allowed to grow under continuous washout conditions, such that weakly bound organisms were selectively lost. Those initially rare founder bacteria expressing a receptor for the immobilized ligand (R+ve) were thus enriched and amplified simultaneously. From an initial culture containing 1 x 10(10) R-ve cells spiked with fewer than 30 R+ve bacteria (<1 in 10(8)), final ratios of R+ve/R-ve bacteria as high as 1 in 12 were observed, representing an enrichment factor of 55 million-fold. This technology has considerable potential for rapid screening of bacterial surface-display libraries and in facilitating directed-evolution studies.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Proteínas da Membrana Bacteriana Externa
/
Proteínas de Bactérias
/
Glicoproteínas de Membrana
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Cromatografia de Afinidade
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Escherichia coli
Tipo de estudo:
Diagnostic_studies
/
Screening_studies
Idioma:
En
Revista:
Biochem J
Ano de publicação:
2001
Tipo de documento:
Article
País de afiliação:
Reino Unido