Microfluidic selection of library elements
A library and element technology, used in chemical libraries, fluid controllers, laboratory containers, etc., to solve problems such as method contamination and result quality deterioration
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example 1
[0051] This example was performed to demonstrate library screening for streptavidin. The microfluidic device has a silicon wafer for the chip and a polydimethylsiloxane (PDMS) substrate. The flow channel has a depth of 20 microns and a width of 60 microns. The receptor contains streptavidin.
[0052] A phage display library encoding dodecapeptides (a phage display library encoding dodecapeptides (M13 phage library from New Zealand Biolabs #E8110S)) was screened against streptavidin immobilized on a PDMS substrate. The microfluidic chip used for this screening is shown in Figures 1 and 2 previously mentioned. 0.1 microgram per milliliter (μg·mL) of streptavidin in phosphate buffered saline (PBS) overnight -1 ) solution to cover the PDMS surface, and streptavidin (provided by the library) was deposited on the PDMS substrate. After washing with PBS, deionized water and 2 After drying under airflow, use 0.5% (weight) bovine serum albumin (BSA) solution and PBS with streptavid...
example 2
[0057] This example was implemented to screen a library of hemagglutinin antigens. A phage display library encoding dodecapeptides (M13 phage library from New Zealand Biolabs #E8110S) was screened against antibody (Ab) targets. This Ab is directed against a synthetic peptide from hemagglutinin influenza virus (9 amino acid sequence YPYDVPYA) and is a monoclonal murine Ab (#H1200-3, IgG, clone 3H428B from USBiological, Ma, USA). The buffer for this library was TBS (tris-buffered saline, ie, 50 mM Tris-HCL, pH 7.5, 150 mM NaCl) with 50% glycerol. The complexity of the library is 2.7×10 19 A transformant. 10 μL of the library contained approximately 55 copies of each sequence.
[0058] The microfluidic device used to screen the library had the same design as the microfluidic device described in Example 1, except that the loading pad used to load the library was replaced with through-holes, which can be obtained from Figure 4 seen in. The via was connected to a micro-port, p...
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