Surface coating for microfluidic devices that incorporate a biopolymer resistant moiety
a technology of biopolymer resistance and microfluidic devices, applied in coatings, thin material processing, transportation and packaging, etc., can solve the problems of loss of separation efficiency, peak tailing, and difficult to achieve the effect of effective suppression of biopolymer adsorption and stable and reproducible electroosmotic flow
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[0184] Example 1 demonstrates that microfluidic devices having a hydroxyethylated poly(ethyleneimine) coating adsorb substantially less protein than analogous microfluidic devices coated with non-hydroxyethylated poly(ethyleneimine).
[0185] Example 2 illustrates enzyme Km and inhibition assays on microfluidic devices that have been coated with hydroxyethylated poly(ethyleneimine). The hydroxyethylated poly(ethyleneimine) surfaces do not significantly adsorb proteins and, thus, eliminate the need for zwitterionic buffer additives.
[0186] Example 3 illustrates the resistance of the coatings of the invention to protein adsorption by assaying bovine serum albumin (BSA) and avidin as representative proteins.
[0187] Example 4 illustrates a synthetic strategy for coating microfluidic devices with a charged poly(ethyleneglycol) derivative wherein indirect covalent bonding via silane or polymer linkers were employed.
[0188] Example 5 illustrates an alternative synthetic strategy for coating ...
example 1
[0192] Example 1 provides a comparison of biopolymer adsorption in devices coated with PEI-1 and PEI-2. PEI-1 is a branched polyethyleneimine with average MW=25,000. PEI-2 is also a branched polyethyleneimine with a base polymer MW=70,000 and has 80% of the amino groups ethoxylated. Both polymers were used to coat microfluidic devices to generate stable, reversed electroosmotic flow. The PEI-2 coating was found to exhibit less protein adsorption than the PEI-1 coating, as judged by comparing electroosmotic flow measurements and PTP1b enzyme assay results for the coated devices.
1.1 Materials and Methods
[0193] 1.1a PEI Coated Devices
[0194] The microfluidic devices used for enzyme assays measuring electroosmotic mobility (EO), had the channel format shown FIGS. 9(a) and 9(b) (with 20 μm deep channels). The top surfaces of new devices were treated with Repel-Silane-ES. The microchannels were cleaned prior to coating by successive rinses with 1N NaOH, water, 1N HCl, water and ethanol...
example 2
[0209] A PTP1B assay using Caliper's microfluidic device is illustrated in Example 2. The microfluidic device surface has been modified with a physically adsorbed high-molecular-mass polyethyleneimine coating. The coating provides a stable, reversed electroosmotic flow surface. Devices coated with 80% ethoxylated polyethyleneimine (PEI-2) exhibited good electroosmotic flow (reversed direction), and show much less protein adsorption for both positively and negatively charged proteins than uncoated glass devices. In uncoated glass devices, the assay is typically run with added sulfobetaine (NDSB) to prevent the adsorption of the enzyme to the walls of the microchannel. Thus, as an additional test of PEI-2 coated devices, the PTP1B assay was run using buffers which did not contain NDSB.
2.1. Materials and Methods
[0210] 2.1a PEI-2 Coated Devices
[0211] The devices used for this assay had a channel format as shown in FIGS. 9(a) and 9(b) (20 μm depth). The channel surfaces of each new d...
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