Method for accurately testing digestion efficiency of proteins in matrix
What is Al technical title?
Al technical title is built by PatSnap Al team. It summarizes the technical point description of the patent document.
A protein enzyme, accurate determination technology, applied in measuring devices, material analysis by electromagnetic means, instruments, etc., can solve the problems of enzyme cleavage efficiency difficult to reach 100%, missed peptide fragments, etc.
Active Publication Date: 2015-04-29
NAT INST OF METROLOGY CHINA
View PDF3 Cites 9 Cited by
Summary
Abstract
Description
Claims
Application Information
AI Technical Summary
This helps you quickly interpret patents by identifying the three key elements:
Problems solved by technology
Method used
Benefits of technology
Problems solved by technology
[0012] Usually, the enzyme cleavage efficiency is calculated as 100%, but in fact, due to the influence of chemical balance and protease inhibitors in the matrix, the enzyme cleavage efficiency is difficult to reach 100%, and there will always be missed cleavage. Peptides
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
Click on the blue label to locate the original text in one second.
Reading with bidirectional positioning of images and text.
Smart Image
Examples
Experimental program
Comparison scheme
Effect test
Embodiment 1
[0124] A method for accurately determining the enzyme cleavage efficiency of a protein in a substrate, characterized in that it comprises the following steps:
[0125] (1) Screen the specific peptides of the target protein, use protease to theoretically digest the target protein, and obtain each theoretically digested peptide; use the Blast tool to search the obtained theoretically digested peptides one by one, and determine the target protein specific enzyme-cut peptides;
[0126] (2) Specific peptides are synthesized from amino acids by chemical synthesis; isotope-labeled specific peptides are synthesized from isotope-labeled amino acids, and the synthesized peptides are purified by reverse-phase high-performance liquid chromatography to obtain pure products , using the isotope dilution mass spectrometry method based on amino acid analysis to determine the mass fraction P of the target peptide in the synthetic peptide peptide ;
[0127] (3) Obtain the pure product of the t...
Embodiment 2
[0176] 1. Instruments used:
[0177] Analytical balance: (METTLER TOLEDO XS205 type, the maximum weighing capacity is 81g, the precision is 0.01mg, the United States); (METTLER TOLEDO XP56 type, the maximum weighing capacity is 52g, the precision is 0.001mg, the United States);
[0178] Constant temperature shaker (S16R-2 type, SHEL LAB, USA);
[0179] Oven (UFE500 type, MEMMERT, the United States);
[0184] 2. Accurate determination of β-lactoglobulin digestion efficiency in milk powder, including the following steps:
[0185] (1) Search the database by means of bioinformatics, and select TPEVDDEALEK (TK-11) as the specific peptide for β-lactoglobulin isotope dilution mass spe...
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
Property
Measurement
Unit
quality score
aaaaa
aaaaa
Login to view more
Abstract
The invention discloses a method for accurately testing the digestion efficiency of proteins in a matrix. The method comprises the following steps: (1) performing specific peptide fragment screening and digestion on target proteins for determining specific digestion peptide fragments; (2) synthesizing the specific peptide fragments; (3) preparing a standard protein mother liquor; (4) performing preliminary measurement on the concentration of the target proteins; (5) performing the digestion and measuring the concentration; (6) adding a diluent, performing the digestion and performing isotopic dilution mass spectrometry measurement on the concentration of the specific peptide fragments; (7) calculating the concentration of the proteins according to the concentration of the specific peptide fragments; (8) calculating the digestion efficiency when the concentration of added target proteins is used in the step (6); (9) repeating the steps (6)-(8), and adding different protein amounts every time to obtain a series of digestion efficiency values; and (10) plotting according to the protein addition amount and the digestion efficiency and extrapolating the digestion efficiency when the added protein amount is zero so as to obtain the accurate digestion efficiency of the proteins. The method has the advantages of high accuracy and good traceability.
Description
technical field [0001] The invention relates to the field of biotechnology, in particular to an accurate method for measuring the enzyme cleavage efficiency of proteins in a substrate. Background technique [0002] With the development of life sciences and the advancement of biotechnology, protein detection has been widely used in clinical testing, food safety, biomedicine and other fields. For example, more than 80% of tumor markers, cardiovascular and cerebrovascular disease markers, and endocrine disease markers are proteins; the detection and labeling of food allergens, and the quality evaluation of recombinant protein drugs are also inseparable from accurate quantification of proteins. In order to ensure the accuracy and comparability of protein detection results in various fields, it is necessary to develop corresponding protein standard substances as the standard for analysis and quantification. Protein standard substances usually use more accurate technical means tha...
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
Application Date:The date an application was filed.
Publication Date:The date a patent or application was officially published.
First Publication Date:The earliest publication date of a patent with the same application number.
Issue Date:Publication date of the patent grant document.
PCT Entry Date:The Entry date of PCT National Phase.
Estimated Expiry Date:The statutory expiry date of a patent right according to the Patent Law, and it is the longest term of protection that the patent right can achieve without the termination of the patent right due to other reasons(Term extension factor has been taken into account ).
Invalid Date:Actual expiry date is based on effective date or publication date of legal transaction data of invalid patent.
Login to view more