Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Rubber-like material for the immobilization of proteins and its use in lighting diagnosis and biocatalysis

A rubber-like, protein-based technology used in biochemical equipment and methods, microbial assay/inspection, luminescent materials, etc.

Inactive Publication Date: 2018-05-11
FUNDACION IMDEA MATERIALES
View PDF4 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0017] The present invention addresses the above-discussed shortcomings and solves the problem of providing new and improved methods of immobilizing and stabilizing a wide range of different proteins, including photoproteins and enzymes, without any need for cross-linking or immobilization

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
  • Rubber-like material for the immobilization of proteins and its use in lighting diagnosis and biocatalysis
  • Rubber-like material for the immobilization of proteins and its use in lighting diagnosis and biocatalysis
  • Rubber-like material for the immobilization of proteins and its use in lighting diagnosis and biocatalysis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0341] Example 1 : Preparation of Rubber-like Materials Containing Immobilized Proteins

[0342] Preparation of proteins and enzymes (expression and purification of recombinant proteins)

[0343] Such as figure 1As shown, several different photoproteins and enzymes were prepared and characterized. Escherichia coli (E.coli) strain M15[pREP4] carrying the appropriate plasmid (both pQE-9 expression constructs containing the N-terminal 6xHis tag from the pQE-9 expression vector, Qiagen) was incubated at 28°C in the presence of Amp (200 μg / ml) and Kan (100 μg / ml) antibiotics grown in LB medium to an optical density at 600 nm of about 0.5. Recombinant protein expression was induced using 1 mM isopropyl-β-D-1-thiogalactoside at 28°C. After 4 hours of growth at 28°C, cells were harvested and frozen at -20°C. Frozen bacterial cells were thawed and lysed chemically using lysozyme and mechanically using a sonicator. Then, following the QIAGEN protocol (Henco K, A handbook for hi...

Embodiment 2

[0357] Example 2 : Preparation of rubber-like materials using different mass ratios of branched polymers and linear polymers and different amounts of buffered aqueous solutions

[0358] The preparation of gels and rubber-like materials according to the present invention uses different mass ratios of branched polymers (in this example, M n 450 Da ethoxylated trimethylolpropane (TMPE)) and linear polymers (in this example, M n 5000 kDa polyethylene oxide (PEO)), as shown in Table 1 and Table 2 below. Rubber formation was performed as described in Example 1.

[0359] Specifically, the above two polymers were mixed in different mass ratios as shown in Table 1 and Table 2 below. Although TMPE is a low-viscosity liquid, PEO does not dissolve even under high-speed stirring conditions. To facilitate this process, different amounts of buffer (again for proteins) were added.

[0360] Table 1:

[0361]

[0362] Table 2:

[0363]

[0364]As summarized in Table 1, until 200 µ...

Embodiment 3

[0369] Example 3: Applications of rubber-like materials containing immobilized proteins as downconversion encapsulation systems

[0370] Figure 5 A schematic diagram showing a hybrid light-emitting diode with a rubber-like material containing immobilized proteins therein as a down-converting packaging system (see Example 1). A commercially available blue emitting LED with an electroluminescence spectrum at 450 nm (from Luxeon) was used in this example. To coat the previous silicone encapsulation in three-dimensional form, the LED can either be immersed in the gel for a few seconds and / or the gel can be deposited by drop coating onto the surface of the carrier. Subsequently, the coating was dried as described in Example 1 above. Devices were driven under constant and / or pulsed current and voltage regimes. In this example, a Keithley 2400 was used to drive the LED at a constant current of 10 mA, and the electroluminescence spectrum and device performance was monitored usin...

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
number average molecular weightaaaaaaaaaa
number average molecular weightaaaaaaaaaa
number average molecular weightaaaaaaaaaa
Login to View More

Abstract

The present invention relates to a process of preparing a rubber-like material containing a protein immobilized therein as well as a corresponding rubber-like material the process comprising the stepsof (a) mixing a protein a branched polymer such as trimethylolpropane ethoxylate and a linear polymer such as poly( ethylene oxide) in an aqueous solution to form a gel and (b) drying the gel to obtain a rubber-like material containing the protein immobilized therein wherein the branched polymer comprises at least three polymeric branches bound to a central branching unit. The rubber-like material allows the immobilization and stabilization of a wide range of different proteins including luminescent proteins as well as enzymes and can particularly advantageously be used as down-converting material for light-emitting diodes (LEDs) for diagnostic applications and in bioreactors.

Description

technical field [0001] The present invention relates to a method for the preparation of rubbery materials containing immobilized proteins therein, and corresponding rubbery materials, said method comprising the steps of: (a) mixing proteins, such as ethoxylated trimethylol A branched chain polymer of propane and a linear polymer such as polyethylene oxide form a gel, and (b) drying the gel to obtain a rubbery material containing immobilized proteins therein, wherein the branched chain polymer Comprising at least three polymeric branches bonded to a central branching unit. This rubbery material can immobilize and stabilize a wide range of different proteins, including photoproteins and enzymes, and is particularly advantageous as a down-converting material for light-emitting diodes (LEDs) for diagnostic applications and in the bioreactor. Background technique [0002] Protein immobilization for biotechnological processes has been considered the most cost-effective means of ...

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
Patent Type & Authority Applications(China)
IPC IPC(8): H01L33/50C07K17/02
CPCB82Y5/00C08G2650/30C09D171/02C07K17/04C09K11/025C09K11/06C12N9/1205C12N9/2431C12N9/92C12N9/96C12Y207/01001C12Y503/01009C08G65/333C07K17/08C08J3/075C08J2371/02C08J2471/02C08J2489/00C09K2211/14C12Q1/008C12Q1/40C12Q1/485C12Q1/533H01L33/56
Inventor R·D·科斯塔,瑞克末U·索尼瓦拉德P·布拉纳,科拓M·D·瓦贝M·普拉塞尔
Owner FUNDACION IMDEA MATERIALES
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com