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

A chemical entity, static technology, applied in the direction of chemical instruments and methods, immobilization on/in organic carriers, preparation methods of peptides, etc.

Inactive Publication Date: 2008-07-30
DRO BIOSYSTEMS SL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, amorphous glasses coated with microfilaments have never been suggested for their use in methods for purifying, separating, detecting, modifying, and / or immobilizing target matrices contained in fluids

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[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.

Embodiment 2

[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....

Embodiment 3

[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(hydr...

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PUM

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Abstract

The present invention discloses a Static Supported Bed (SSB) for purifying, isolating, detecting, modifying, and / or immobilizing a chemical entity of interest or a biological entity of interest present in a fluid. The static support bed of the present invention comprises one or more microwire supports suitable for attachment of a chemical entity of interest or a biological entity of interest.

Description

technical field [0001] The present invention relates to a statics support bed for purifying, separating, detecting, modifying, and / or immobilizing a target chemical entity or target biological entity present in a fluid. Background technique [0002] Various analytical, biochemical and diagnostic methods involve the immobilization of specific reagents or biological binding partners of biomolecules to high surface area matrices. [0003] On the one hand, cells are usually cultured in reactors to manufacture biological and pharmaceutical products. Such cells may be animal, plant, fungal or microbial cells. To maintain cell cultures, in general, oxygen and other nutrients must be supplied to the cells. Cell cultures are typically maintained within the reactor by perfusion, wherein cell culture medium including oxygen and other nutrients is passed through the cell culture reactor. However, cell culture reactors can only support small cell loads per unit of reactor volume. The...

Claims

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Application Information

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IPC IPC(8): C12N11/04C12N15/10C07K17/14C07K1/14C03B37/026H01B3/08B01L99/00
CPCC03B37/026C12N11/04Y02P40/57
Inventor S·马科斯A·卡斯特鲁伊斯I·洛伊纳斯J·A·蓬波索V·茹科娃
Owner DRO BIOSYSTEMS SL
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