Elution Dynamics of M13 Bacteriophage Bound to Streptavidin Immobilized in a Microfluidic Channel

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Microfluidic biopanning can reduce a sample size and improve the reproducibility of biopanning. However, there is no practical guideline for optimal experimental conditions to identify high affinity peptides, including incubation and washing times and washing and elution flow rates. Here we investigated the elution dynamics of M13 phage displaying a well-known streptavidin-binding peptide motif, histidine-proline-glutamine (HPQ), to streptavidin immobilized in a polydimethylsiloxane microfluidic channel via a biotin-polyethylene glycol linker. The phages bound to streptavidin were eluted using an acidic buffer, which weakens the molecular interactions of HPQ and streptavidin. Various washing flow rates were examined to determine the effects of drag forces on the elution of phages, and the total force per phage combined with the drag force was calculated. The critical washing drag force and washing time were 5.9 x 10(-8) dyne and 1.1 min, respectively, as obtained by fitting the data using a sigmoidal function. The dissociation rate constant of HPQ-displaying phage (2.0 x 10(-2) min(-1)) was also obtained from the double exponential decay fitting. Our work provides a practically useful approach to the optimization of experimental parameters for the design of a microfluidic biopanning platform.
Publisher
KOREAN BIOCHIP SOCIETY-KBCS
Issue Date
2016-03
Language
English
Article Type
Article
Keywords

PHAGE DISPLAY; OPTIMIZATION; FABRICATION; ANTIBODIES; SELECTION; PEPTIDES; SURFACES; FORCE; FLOW

Citation

BIOCHIP JOURNAL, v.10, no.1, pp.48 - 55

ISSN
1976-0280
DOI
10.1007/s13206-016-0107-x
URI
http://hdl.handle.net/10203/208670
Appears in Collection
NT-Journal Papers(저널논문)MS-Journal Papers(저널논문)
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