Static support bed for purification, separation, detection, modification and/or immobilization of target entities and method using thereof

[0109] Example 1: Example of microwire support fabrication

[0110] Fabrication of a continuous microwire support with an external diameter of 24.4 μm is described below:

[0111]Glass tubes having an external diameter of 7 to 10 mm and a wall thickness of 1.0 to 1.4 mm, filled with a metal alloy consisting of 69% cobalt, 4% iron, 1% nickel, 13% boron and 11% silicon, 0.9 to 1.5 kg The feed rate between centimeters per minute is fed into the induction oven of the microfilament fabrication machine. The oven temperature is set between 1260 and 1330°C. The resulting metal-filled microwire supports were cooled in running water and wound at a winding speed between 150 and 250 m/min to form spools, which were stored at room temperature until use.

[0112] The thickness of the glass layer of the microwire support, as well as the overall diameter of the microwire support, can be modified by adjusting the temperature of the induction oven, winding speed and feeding speed.

[0113] Example 2: Fabrication of EcoR I-Activated Microfilament Supports (C-S Bonds)

[0114] The microwire support was treated with a mixture of 7 volumes of sulfuric acid and 3 volumes of 30% hydrogen peroxide at room temperature for 30 minutes. The support is then rinsed thoroughly in running water, then in ethanol, and then in chloroform. Finally, the support was dried under nitrogen flow. Next, the microwire support was treated with 2% (3-aminopropyl)triethoxysilane in water under a nitrogen atmosphere at room temperature. The support was then rinsed in dichloromethane and exposed to a flow of nitrogen. After the ethanol wash, the microwire support was treated with 2 mM 4-maleimidobutyrate N-hydroxysuccinimidyl ester in ethanol for 16 hours and rinsed in ethanol. Then the microfilament support was treated with 600 units of EcoR I enzyme/m microfilament support in TE buffer (0.1M tris(hydroxymethyl)aminomethane and 1mM ethylenediaminetetraacetic acid in water at pH 8.0) ; pH adjusted to 8.0 with hydrochloric acid) for 16 hours and washed in TE buffer.

[0115] Example 3: Application of EcoR I-Activated Microfilament Supports to EcoR I Sensitive Plasmids processing

[0116] A 2.5 μg/ml pCMS-EGFP plasmid (BD Biosciences, cat. no. 6101-1 ) containing a unique target site for the EcoR I enzyme was added in the presence of 50 mM sodium chloride, 100 mM tris(hydroxymethyl)aminomethane, 10 mM magnesium chloride and 0.25% Triton X-100 in water (pH 7.5) were exposed to the activity of the EcoR I-activated microwire support at 37°C for 4 hours. The activity of EcoR I-activated microwire supports on plasmid molecules was analyzed by agarose gel electrophoresis. Non-activated actin support was used as negative control group.

[0117] Electrophoretic analysis of the activity of EcoR I-activated microfilament supports on plasmid molecules:

[0118] A 600 μl sample of the supernatant obtained after treatment with EcoR I-activated actin support was precipitated in 70% ethanol, dissolved in water, and dissolved in TAE (0.04M tris(hydroxymethyl)aminomethane 0.001M ethylenediaminetetraacetic acid, pH adjusted to 8.5 with glacial acetic acid) 0.8% agarose colloid, using TAE as the gel running buffer, electrophoresis at 10.5 volts/cm for 40 minutes. Colloids were stained with 0.5 μg/ml ethidium bromide in TAE for 20 minutes and visualized under UV light. Only the EcoR I-activated microfilament support had any effect on the plasmid molecule.