Biocompatibility coating for continuous analyte measurement
A biocompatible, analyte-based technology applied in the field of biosensors
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Embodiment 1
[0157] Example 1: Protein adsorption
[0158] Protein adsorption to polymer layers was experimentally studied with a quartz crystal microbalance (QCM) using a Q-Sense QSX301 device. In these experiments, a QCM substrate (Q-Sense) was first coated with a polymer or polymer network. The coated sample is placed in the measuring cell and induced to vibrate. Record the resonance frequency. Subsequently, the sample surface was covered with PBS buffer solution, and the polymer coating started to swell until it reached an equilibrium state, whereby the resonance frequency did not change any more. Subsequently, start the protein adsorption experiment by recording the vibrational frequency again, recording the final resonance frequency in buffer solution. After about 15 minutes, the sample surface was further covered with buffer solution, the protein solution was added to the measurement cell, and allowed to cover the sample surface for about 15 minutes, after which the surface was w...
Embodiment 2
[0160] Example 2: Cell Adhesion
[0161] Cell adhesion was tested on glass coverslips as substrates. The coverslip was constructed of quartz glass and had a circular shape with a diameter of 1.5 cm (VWR‚ ECN 631-1579). Between steps in the experiment, place coverslips individually in 24-well plates with the coating side up and store in a low-dust and low-bacteria environment. Handle the coverslip with vacuum tweezers. Coat the surface of the cover glass directly with p(BUMA / HEMA / HPMA) via spin coating; for p(DMAA) coating, first coat the cover glass surface with polystyrene (PS) by spin coating, followed by coating with p(DMAA) coating followed by UV irradiation (365 nm, 0.35W / m 2 ,2 minutes). Uncoated and PS-coated coverslips were used as controls.
[0162] To avoid microbial contamination of the cells, isolate the cultured cells by immersing them in 70% ethanol / water or Perform (25% ethanol, 35% propan-1-ol, and 40% water) for 15 s and by handling coverslips in a steril...
Embodiment 3
[0165] Embodiment 3: toxicity test
[0166] For toxicity testing, both sides of PET substrates (2×2 cm) were network-coated with p(DMAA) by spin coating with p(DMAA), followed by UV irradiation. Samples were sterilized by electron beam (25 kGy) before use. Three test samples were submerged in 4 ml of cell culture medium for 24 hours at 37°C. 1 ml of the incubated cell culture medium (eluate) was transferred to the wells of a fresh 96-well plate, L929 fibroblasts were added and incubated for another 24 h, and the viability of the cells was subsequently measured using the XTT-assay. Results are shown as percent cell growth, using untreated cell culture medium as a reference (eg, 100%). Therefore, the lower the percentage growth value, the more toxic the sample is in the cells. Fragments cut from latex gloves and eluates prepared as described above from p(BUMA / HEMA / HPMA) coated PET substrates were used as controls. For example, pieces cut from latex gloves were used as positi...
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