The use of affinity-coated surfaces to immobilize cells is commonly applied to cellular separations and microfluidic devices. Most of the time, these devices are discarded after a single use and the time and cost of replacing them can rapidly add up. The goal of this study was to assess the reusability of affinity coated microfluidic devices previously used to capture Jurkat T-lymphocytes for real-time analysis. The affinity surface is created using a series of proteins that result in a layer of anti-CD71 which can bind and capture Jurkat cells. The reusability of these microdevices was assessed by measuring the binding efficiency (number of cells bound post-wash/number of cells bound pre-wash) after cleaning the devices with various reagents. Results indicate that devices cleaned with ethanol after being used to capture cells can be recoated with the affinity surface proteins and maintain a high binding efficiency. The average binding efficiency for this device was 95.9% ± 4.3% before cleaning and 95.5% ± 2.1% after cleaning with ethanol and reapplication of the affinity surface. Devices cleaned with ethanol but not recoated with the affinity surface showed an initial binding efficiency of 96.2% ± 3.3% that dropped to 24.8% ± 14.8% after cleaning with ethanol. The results indicate that ethanol significantly reduces the affinity of the surface for the cells. Further investigation of other cleaning reagents, such as NaOH and PBS (as a control), will be discussed. Lastly, all experiments conducted with anti-CD71 will be repeated using anti-CD95 for cell capture. Overall, the results indicate that affinity surfaces can be cleaned and reused or potentially reused without cleaning if a decrease in binding efficiency is acceptable for the experiment. Adaptation of the methods presented here could save money and time for research involving affinity coated microfluidic devices in the future.

Investigation of the reusability of affinity surfaces for cell capture and analysis

255th National Meeting (2018)

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