Biosensor

Inactive Publication Date: 2009-09-24
CCI HLDG INC +1
View PDF10 Cites 15 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0009]Recently, the number of patients of hyperlipemia, which is a lifestyle-related disease, has been growing steadily, and thus it is a present state that development of a sensor, which is capable of measuring, quickly and in high accuracy, concentration of neutral fat contained in a sample like blood or the like, is strongly desired.
[0010]In these circumstances, it is an object of the present invention to provide a sensor, which is capable of measuring, quickly and in high accuracy, concentration of neutral fat from a sample like a biological sample or the like, without executing pretreatment of the sample.
[0011]The present inventors have found that, in a biosensor for measuring concentration of neutral fat, based on value of current flowing in the electrode system, having: an insulating substrate; an electrode system having a working electrode and a counter electrode, formed onto the insulating substrate; and a reaction layer formed at the upper part or the vicinity of the electrode system; by inclusion of a lipoprotein lipase, a glycerol dehydrogenase and an electron mediator in the reaction layer, concentration of glycerol in a sample can be measured rapidly and in high accuracy, and have thus completed the present invention.

Problems solved by technology

However, a triglyceride sensor described in the above JP-A-59-210356 measures fatty acids liberated by a glycerol ester hydrolase, using a pH sensitive, ion selective electric field effect transistor, and because response value is proportional to logarithm of sample concentration, measurement accuracy is insufficient.
And this sensor also has a problem that an apparatus becomes complicated.
Since the sensor requires supply of a measurement sample from its outside, it requires a large quantity of samples, thus makes measurement difficult using a small amount of a sample.
Further, a sensor described in the above JP-A-2001-343349, is configured by different layers for an enzyme and an electron mediator, which makes an apparatus extremely complicated, and requires use of two kinds of expensive enzymes, a glycerol kinase and a glycerophosphate oxidase, as well as a lipoprotein lipase.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Biosensor
  • Biosensor
  • Biosensor

Examples

Experimental program
Comparison scheme
Effect test

reference example 1

[0071]Into a 500-mL Sakaguchi flask, 400 mL of culture media composed of sorbitol (2% by weight), yeast extract (0.3% by weight), meat extract (0.3% by weight), corn steep liquor (0.3% by weight), polypeptone (1% by weight), urea (0.1% by weight), KH2PO4 (0.1% by weight), MgSO4.7H2O (0.02% by weight), CaCl2 (0.1% by weight), with a pH of 7.0, was transferred, by 100 mL per one flask, and autoclaved at 121° C. for 20 minutes.

[0072]As an inoculum, a strain of Gluconobacter Oxydans NBRC 3291, was inoculated by one platinum loop and cultured at 30° C. for 24 hours to make a seed culture media.

[0073]Then, 6.6 L of the culture media prepared with the same composition as in the above, was transferred to a 10-L jar fermenter, and autoclaved at 121° C. for 20 minutes, followed by gradually cooling and subsequently transferring of a 400 mL of the seed culture media, which was cultured under 750 rpm and an airflow amount of 7 L / minute, at 30° C. for 24 hours.

[0074]The culture media was subject...

example 1-1

[0075]As a sensor substrate formed with an electrode system, a commercial sensor electrode (manufactured by BVT Co., Ltd; ACl.W5.R1) was prepared. In this sensor substrate, the electrode system has a diameter of about 6 mm, and nearly circular shape. Furthermore, at the upper part of the electrode system prepared above, a PET sheet, with a thickness of 0.2 mm, and having a hole with a diameter of 6 mm, was glued together to form a spacer. Into the inside of the hole of the spacer on the electrode system, 10 μL of phosphate buffered saline mixed with GlyDH (20 U / mL) obtained in the above Reference Example 1, the lipoprotein lipase (manufactured by Amano Enzyme Inc., 10,000 U / mL), and 1-methoxy-5-methylphenazinium methylsulfate (manufactured by DOJINDO Laboratories, 10 mmol / L; hereinafter referred to as “m-PMS”) as an electron mediator, was dropped, and dried at room temperature, to form a reaction layer.

[0076]Onto this reaction layer, 10 μL of a sample liquid containing 50 mg / dL of t...

example 1-2

[0081]After dropping an aqueous solution of 0.5% (w / v) of sodium carboxymethyl cellulose (manufactured by Wako Pure Chemical Industries Ltd; hereafter may be referred to as “CMC”) onto the electrode system used in Example 1-1, and drying, a 0.5% CMC solution containing GlyDH (20 U / mL) obtained similarly as in Example 1-1, the lipoprotein lipase (10,000 u / mL) and m-PMS (10 mmol / L) was dropped, and dried to form a reaction layer.

[0082]Correlativity between response current value and triolein concentration was studied similarly as in Example 1-1, by using the biosensor. The result is shown in FIG. 5.

[0083]As shown in FIG. 5, although the resultant response current value was reduced a little by effect of CMC, higher correlativity between response current value and concentration of neutral fat was observed as compared with Example 1.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
Lengthaaaaaaaaaa
Electrical resistanceaaaaaaaaaa
Electrical resistanceaaaaaaaaaa
Login to view more

Abstract

It is an object of the present invention to provide a sensor, which is capable of measuring, quickly and in high accuracy, concentration of neutral fat from a sample such as a biological sample or the like, without executing pretreatment of the sample. This object is attained by a biosensor for measuring concentration of neutral fat, based on value of current flowing in the electrode system, having: an insulating substrate; an electrode system having a working electrode and a counter electrode, formed onto the insulating substrate; and a reaction layer having a lipoprotein lipase, a glycerol dehydrogenase and an electron mediator, formed at the upper part or the vicinity of the electrode system.

Description

TECHNICAL FIELD[0001]The present invention relates to a biosensor, which is capable of determining, quickly and in high accuracy, quantity of neutral fat contained in a biological sample or the like.BACKGROUND ART[0002]In general, as a measurement method for neutral fat, there are (1) a method using chromotropic acid, (2) an acetylacetone method, (3) an enzyme method, and (4) a method using a nephelometer, however, any of these methods requires complicated analysis operations and long analysis time. Therefore, a biosensor has been developed, as a method for determining, simply and quickly, quantity of objective substances without execution of dilution or stirring of a sample liquid, in measurement of specific components contained in the sample.[0003]As such a biosensor for measurement of neutral fat, there is a triglyceride sensor characterized in that a glycerol ester hydrolase is immobilize onto a porous polymer membrane or a hydrophilic uniform polymer membrane covered onto a gat...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): G01N27/26G01N27/327
CPCC12Q1/004C12Q1/61C12Q1/005C12N11/096G01N27/327G01N33/48G01N27/3272
Inventor MURASE, HIRONOBUKUWAHARA, MOTOAKIYAMADA, MASAYUKI
Owner CCI HLDG INC
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap