A kind of protein A adsorption material for targeted adsorption and preparation method thereof
An adsorption material and protein technology, which is applied in chemical instruments and methods, suction devices, and other chemical processes, can solve the problems of short coupling spacer arms, high protein A shedding, and low adsorption capacity, so as to improve adsorption performance, High adsorption efficiency and low shedding
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Examples
Embodiment 1
[0043] The synthesis of embodiment 1 epoxy-activated agarose gel
[0044] After washing 100g of agarose gel (Sepharose 6FF) with about 10 times the volume of water for injection, add 100mL of 1M NaOH solution, 30mL of epibromohydrin and 70mL of 1,4-butanediol diglycidyl ether and 0.2 g sodium borohydride, stirred, and reacted at 30° C. for 1.5 h. After the reaction, rinsed with a large amount of water for injection to obtain an epoxy-activated agarose gel.
Embodiment 2
[0045] Synthesis of Example 2 Protein A Adsorption Material
[0046] In a 500mL reaction vessel, add 100mL of epoxy-activated agarose gel synthesized in Example 1, 150mL of borate buffer solution of 0.1mol / L, control the pH value of the system from 7.5 to 8.5, add 14g of genetically engineered recombinant protein A, React in a constant temperature system at 37°C for 20h, stop the reaction, rinse the packing with about 10 times the volume of water for injection, after rinsing, add 200mL of 0.2M ethanolamine solution to block unreacted epoxy groups, and react at 20°C for 10h. After the reaction, rinse with a large amount of water for injection, and finally store it in a solution containing preservatives.
Embodiment 3
[0047] Embodiment 3 adsorption performance contrast
[0048] Take respectively the protein A adsorption material synthesized in Example 2, the protein A adsorption material synthesized using diamine and dialdehyde (the only difference from Example 2) as the spacer arm, and the use of periodate (the only difference from Example 2) ) Each 1mL protein A adsorption material prepared by oxidizing and activating agarose-coupled protein A was installed in a Ф0.8×5cm column, and the column was fully washed with normal saline, and 10mL plasma was passed through the column at a flow rate of 1mL / min. I to G, adsorbed on the adsorption material. Then rinse the plasma with 30 mL of normal saline, use 0.01M citric acid-disodium hydrogen phosphate buffer solution with pH=2.8 as the eluent to elute the human immunoglobulin IgG on the adsorption material, and collect the eluted solution, the eluent was fixed to 100mL, and the absorption peak absorption value of the eluent near 280nm was detec...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
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