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Sensor for detection of carbohydrate

By using sensors based on competitive binding of energy transfer donor-acceptor pairs to detect substances and animal lectins in diabetic patients, stable monitoring of glucose levels is achieved, which solves the pain and discomfort caused by frequent blood collection in the existing technology. stability issues and improve the reliability and accuracy of monitoring.

Inactive Publication Date: 2008-01-16
PRECISENSE AS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these lectins are estimated to have similar disadvantages as Con A

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0280] Example 1: Staining of MRL

[0281] Change human MBL buffer to (by dialysis) containing 150 mM NaCl and 1.25 mM Ca 2+ , 10mM NaHCO at pH 8.7 3 . The dye used for staining is AlexaFluor TM 594 succinimidyl ester (AF594-SE) (Molecular Probes, Eugene, Oregon, USA). The dye was dissolved in dry DMSO and added slowly (10 minutes) to MBL in bicarbonate buffer. The reaction was allowed to proceed for 1 h. Staining was performed with a 15-fold molar excess (relative to the polypeptide unit) of the dye. By using 10mM Tris buffer at pH 7.4, 150mM NaCl and 1.25mM Ca 2+ Purify by dialysis. The degree of labeling of the stained protein as determined by UV spectroscopy was 2.3 dyes per MBL subunit.

Embodiment 2

[0282] Embodiment 2: Preparation of aminodextran

[0283] 150 kDa dextran (6.00 g, 0.4 μmol) was dissolved in 250 mM K at pH 11.5 2 HPO 4 (600mL). Sodium borohydride (3 g, 0.08 mol) was added followed by divinyl sulfone (15 ml, 0.15 mol). The reaction mixture was stirred at RT for 30 minutes before being neutralized to pH 7.2 with concentrated hydrochloric acid. After stirring for 30 minutes, the resulting mixture was dialyzed (MWCO 10-12 kDa) against water (3 x 25 L). The contents were transferred to an Erlenmeyer flask and 24% aqueous ammonia (200 mL) was added. After 2h, the pH was adjusted to 10.5 and the reaction was stirred overnight. Excess ammonia was removed by dialysis (MWCO 10-12 kDa) in water (8 x 25 L), and the entire contents were lyophilized to give aminodextran 5.75 g (78%, based on 185 kDa MW) as a white fluffy substance. aminodextran). Rough estimates of molecular weight, amine incorporation, and amount of divinylsulfone incorporated were made using ...

Embodiment 3

[0284] Embodiment 3: the preparation of hexamethoxy-crystal violet succinimide ester (HMCV-1)

[0285] Synthesis of HMCV-1:

[0286]

[0287] Synthetic route 1: I) 4-(N-methylamino)-butyric acid hydrochloride (1 equivalent), diisopropylethylamine, in acetonitrile, 20°C, 20h. II) Dimethylamine (excess). III) TSTU, diisopropylethylamine, in acetonitrile, 20°C, 2h.

[0288] 4a (BF4 - ): 4-(Methylamino)butyrate hydrochloride (1.36 g; 8.8 mmol), 1 (5.0 g; 8.3 mmol), and diisopropylethylamine (5 mL) were dissolved in acetonitrile (120 mL). The reaction mixture was stirred at 30-35 °C for 22 h under a dry nitrogen atmosphere. Aqueous dimethylamine (40 mL of a 40% solution) was added and the reaction mixture was stirred for an additional four days. The solvent and excess dimethylamine were removed under vacuum, and the remaining material was dissolved in chloroform. The chloroform solution was washed twice with brine and washed with MgSO 4 dry, then evaporate the solvent a...

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PUM

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Abstract

A sensor for the detection or measurement of carbohydrate analyte (such as glucose) in fluid comprises components of a competitive binding assay the readout of which is a detectable or measurable optical signal (such as a FRET assay) retained by a material that permits diffusion of analyte but not the assay components, the assay components comprising: an animal lectin; and an analyte analogue capable of competing with analyte for binding to the lectin.

Description

technical field [0001] The present invention relates to sensors, methods of making sensors and methods of using sensors. [0002] The sensor can be used to measure or monitor sugars in fluids, such as glucose in bodily fluids, using optical techniques. [0003] The sensor is particularly suitable for situations where glucose levels have to be monitored precisely and / or glucose has to be measured repeatedly, eg in diabetes monitoring. Background technique [0004] In diabetes monitoring, routine measurement of glucose in the blood is necessary to ensure proper insulin dosing. Additionally, it has been shown that better control of blood sugar levels in the long-term care of diabetic patients can delay, if not prevent, the onset of retinopathy, circulatory problems and other degenerative diseases associated with diabetes. Therefore, diabetic patients need reliable and precise self-monitoring of blood glucose levels. [0005] It is desirable to measure blood glucose in the ra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543
CPCC08B37/0063A61B5/145G01N33/66G01N33/54373A61B5/14532
Owner PRECISENSE AS
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