cp4-epsps monoclonal antibody and preparation method thereof

A CP4-EPSPS and monoclonal antibody technology, which is applied in the field of preparation of CP4-EPSPS monoclonal antibody and CP4-EPSPS monoclonal antibody, can solve the problems of limited monoclonal antibody preparation, high cost, and limited demand for antibodies

Active Publication Date: 2020-10-30
杭州贤至生物科技有限公司
View PDF6 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the small amount of ascites produced by a single mouse and the large individual differences, the obtained anti-glyphosate gene protein CP4-EPSPS monoclonal antibody has large differences between batches. Discrepancy issues make detection less accurate
In order to solve the above problems, another method is to prepare antibodies through CHO cell expression, which can avoid the use of a large number of mice and ensure that the differences between different batches of antibodies are small, but the demand for antibodies is limited, and cell expression is continuous. The cost of artificially controlling cell expression of antibodies is high, which severely limits the preparation of monoclonal antibodies

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
  • cp4-epsps monoclonal antibody and preparation method thereof
  • cp4-epsps monoclonal antibody and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 Preparation of CP4-EPSPS Monoclonal Antibody

[0052] a. Total RNA extraction

[0053] CP4-EPSPS monoclonal cells: from Hangzhou Xianzhi Biotechnology Co., Ltd.

[0054] Preparation method: Take 2 mL of cell suspension, centrifuge to remove the supernatant, collect CP4-EPSPS monoclonal cells in the logarithmic growth phase, add 1 mL of TRIzol reagent to the cell pellet, and let it stand for 5 minutes. Add 200 μL of chloroform, shake vigorously for 15 seconds, let stand at room temperature for 3 minutes, and then centrifuge at 12000 r / min for 15 minutes. Transfer the upper water sample layer to a new centrifuge tube, add 0.5mL isopropanol, and let stand at room temperature for 10min. Then centrifuge at 12000r / min for 10min. Discard the supernatant, add 1mL 75% ethanol, centrifuge at 7500r / min for 5min, dry the precipitate, add 50μL RNase-free ddH 2 O. Its purity was measured and quantified with an ultraviolet spectrophotometer. The OD260 / OD280 was 1.86, wh...

Embodiment 2

[0076] Embodiment 2: the preparation of colloidal gold immunochromatography test strip

[0077]S01: Take 10ml of 0.01% colloidal gold solution, add 20uL of 0.2mol / L potassium carbonate solution, mix thoroughly, add 100ug CP4-EPSPS monoclonal antibody, react at room temperature for 2 hours, add 1ml of 10% bovine serum albumin (BSA), and block After 2 hours, centrifuge (7500rpm / min, 20 minutes), discard the supernatant, and dissolve the precipitate with 1ml of reconstitution solution, and spray it evenly on the Glass fiber (width 6mm), and then placed in an electric heating constant temperature incubator (Shanghai Yiheng Scientific Instrument Co., Ltd.) at 37°C for 30 minutes.

[0078] The relevant solution formula is as follows:

[0079] 0.01% colloidal gold solution: 1ml of 1% chloroauric acid solution, 1.4ml of 1% citric acid solution, add ultrapure water to dissolve the reaction and make the volume to 100ml.

[0080] 1% chloroauric acid solution: 1g AuCL3.HCl.4H2O powder i...

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 provides a CP4-EPSPS monoclonal antibody and belongs to the technical field of bioengineering. Existing problems of high preparation cost of CP4-EPSPS monoclonal antibody and the like are solved. The CP4-EPSPS monoclonal antibody comprises a heavy chain and a light chain, wherein the nucleotide sequence of the heavy chain is shown in a sequence table (1), and the nucleotide sequenceof the heavy chain can encode the amino acid sequence of the heavy chain shown in a sequence table (2). The light chain nucleotide sequence is shown in a sequence table (3) and can encode the light chain amino acid sequence shown in a sequence table (4). The CP4-EPSPS monoclonal antibody has the advantages of low cost and the like.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a CP4-EPSPS monoclonal antibody and a preparation method of the CP4-EPSPS monoclonal antibody. Background technique [0002] Glyphosate (N-phosphonomethyl glycine) is a broad-spectrum herbicide that can control a variety of annual and perennial monocotyledonous or dicotyledonous weeds and herbaceous plants. , low dosage per unit area, easily degraded in the soil environment and harmless to the environment, etc., glyphosate has become a herbicide with the largest usage and the largest promotion area in the world. Although glyphosate is the best choice as a herbicide, because it is a non-selective herbicide, it will cause harm to weeds and crops, affecting the growth and yield of crops. For this reason, it has become the mainstream of research to introduce glyphosate-resistant CP4-EPSPS gene through transgenic technology into the plant genome to form glyphosate-resistant tr...

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 Patents(China)
IPC IPC(8): C07K16/16C12N15/85C12N15/66C12N15/10
Inventor 胡祥叶曹丹琴朱伟刘清泉项美华武戌青
Owner 杭州贤至生物科技有限公司
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