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

Method for detecting activity of residual trypsin in cell product

A trypsin and detection method technology, applied in the field of protein detection, can solve the problems of unsuitable detection of trypsin activity, etc., and achieve the effects of simple method, improved sensitivity, and increased reaction temperature

Inactive Publication Date: 2017-01-04
TIANJIN AMCELLGENE ENG
View PDF0 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the commonly used methods are all for trypsin preparations with relatively high trypsin content, and are not suitable for detecting trace amounts of trypsin activity.

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
  • Method for detecting activity of residual trypsin in cell product
  • Method for detecting activity of residual trypsin in cell product
  • Method for detecting activity of residual trypsin in cell product

Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment

[0023] A method for detecting residual trypsin activity in cell products, comprising the following steps:

[0024] (1) Blank experiment: Take 2.5-3.5ml of the substrate solution, add 150-250μl of 0.001mol / L hydrochloric acid solution, mix well, and use it as a blank experiment.

[0025] (2) Determination of residual trypsin activity of cell products: take 100 μl of cell product solution, add 2.5-3.5 ml of substrate solution and 80-120 μl of phosphate buffer solution, immediately time and mix evenly, and measure the absorbance OD value at 253nm A2, while keeping the reaction system at 36-38°C; then measure the absorbance OD value A1 at 253nm at the point of reaction for 25 minutes; A1-A2 represent the residual trypsin activity in the product.

[0026] In order to further optimize the above technical scheme, take 3.0ml of the substrate solution, add 200μl of 0.001mol / L hydrochloric acid solution, mix well, and use it as a blank experiment; the blank experiment is used to offset ...

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

No PUM Login to View More

Abstract

The invention discloses a method for detecting the activity of residual trypsin in a cell product. The method is characterized in that trypsin is used to catalyze hydrolysis of N-benzoyl-L-arginine ethyl ester hydrochloride into N-benzoyl-L-arginine hydrochloride and ethanol, and detecting the absorbance of the hydrolyzed reaction system at a wavelength of 253 nm in order to determine the activity of trypsin. The new trypsin activity detection method is obtained when the optimum reaction temperature is 37 DEG C and the optimum detection time is 25 min after the reaction starts; and the method has high sensitivity, so the method can be used to detect the activity of the residual trypsin in the cell product. The method for detecting the activity of residual trypsin in the cell product is simple, sensitive and practical.

Description

technical field [0001] The invention relates to the technical field of protein detection, in particular to a method for detecting residual trypsin activity in cell products. Background technique [0002] Trypsin is an essential reagent in the cell culture process and is used for the digestion of adherent cells. However, when the cell products are used in the human body, the residual trypsin activity may cause potential harm to the user. Therefore, residual trypsin activity in cell preparations should be detected and limited. [0003] As a proteolytic enzyme, trypsin has high specificity for the bonds formed by the carboxyl groups of basic amino acids (such as arginine and lysine) and the amino groups of other amino acids; The amide bond or ester bond formed by the carboxyl group; therefore, the activity of trypsin can be determined by using the amide or ester compound containing these bonds as a substrate. Currently commonly used methods are aimed at trypsin preparations w...

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): C12Q1/37C12Q1/02
Inventor 韩忠朝王斌冯杰韩之波耿洁
Owner TIANJIN AMCELLGENE ENG
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