Sulfadimidine affinity membrane and application of sulfadimidine affinity membrane in antibody separation and purification

A sulfamethazine, separation and purification technology, applied in the field of biomedical materials, can solve the problems of low purity, small processing capacity, long time, etc., and achieve high purity and activity, mild operating conditions, and high selectivity. Effect

Active Publication Date: 2012-07-11
MABPLEX INT LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the commonly used methods for separating and purifying antibodies include salting-out method, octanoic acid-saturated ammonium sulfate method, affinity chromatography, gel filtration chromatography, hydrophobic chromatography, and ion-exchange chromatography, etc., but the first two Various methods can only obtain antibodies with low purity; although affinity chromatography has good selectivity, there are time-consuming steps such as column loading and pretreatment, and the processing volume is small; gel filtration chromatography, hydrophobic chromatography, As well as ion exchange chromatography, there are also problems such as small processing capacity, long time, and column packing.

Method used

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  • Sulfadimidine affinity membrane and application of sulfadimidine affinity membrane in antibody separation and purification
  • Sulfadimidine affinity membrane and application of sulfadimidine affinity membrane in antibody separation and purification
  • Sulfadimidine affinity membrane and application of sulfadimidine affinity membrane in antibody separation and purification

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Example 1: Preparation of Sulfamethazine Affinity Membrane

[0030] (1) Disperse 10 pieces of cellulose acetate membranes (Hangzhou Kenuo, Φ=10cm) in 100mL NaOH solution with a concentration of 2mol / L, and hydrolyze it in a water bath vibrating shaker (175rpm) at 70°C for 1.5h. After cooling down to 40°C, add a solution of epichlorohydrin with a volume fraction of 20%, and react at 40°C for 4 hours. After the reaction, wash with deionized water to neutral under suction filtration conditions to obtain an epoxy-activated membrane.

[0031] (2) Dissolve 1.0 g of sulfamethazine in a PBS buffer solution of pH 10.5, 20 mmol / L, and disperse 10 pieces (about 5 g) of the epoxy-activated film obtained in step (1) in the buffer solution , shake (175rpm) in a reaction water bath at 55°C for 36h. Then wash with deionized water under the condition of suction filtration until the ultraviolet absorption of the filtrate at 280nm is close to 0, and after drying, the sulfamethazine affin...

Embodiment 2

[0034] Example 2: Application of Sulfamethazine Affinity Membrane in Antibody Separation and Purification

[0035] Get the sulfamethazine affinity membrane 10~15 that embodiment 1 makes, it is packed in the film cup (such as figure 1 ), dilute 10ml of ascites with pH 5.5, 20mmol / L PBS buffer solution to 100mL, and then use a peristaltic pump to pump it into the membrane cup at a flow rate of 2ml / min to allow it to fully adsorb with the ligand on the membrane. Deionized water was pumped in at a flow rate until the filtrate had near zero UV absorbance at 280 nm. Then the elution buffer (PBS of 20 mmol / L at pH 5.5+1.5 mol / L NaCl) was pumped in at the same flow rate. The eluted substances were collected, centrifuged at 3000rpm for 15min with an ultrafiltration centrifuge tube, then distributed into 1.5mL tubes, and frozen for preservation.

[0036] The purity of the antibody was detected by SDS-PAGE, and the results were as follows image 3 As shown, 1 indicates protein marker,...

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Abstract

The invention discloses a sulfadimidine affinity membrane; a preparation method of the sulfadimidine affinity membrane comprises the following steps: (1) taking a cellulose acetate membrane, completely hydrolyzing under an alkaling condition, then adding epoxy chloropropane to perform epoxy activation, and then using deionized water to wash to be neutral after reaction under a suction filter condition, so as to obtain the epoxy activated membrane; (2) dissolving the sulfadimidine into an alkaline buffer solution with pH of 10-12, then dispersing the epoxy activated membrane obtained in the step (1) in the above-mentioned solution, reacting at the temperature of 50-65 DEG C for 24h-36h, then using deionized water to wash under the suction filter condition to make UV absorption of a percolate at 280nm close to zero, and then drying the solution to get the sulfadimidine affinity membrane. The sulfadimidine affinity membrane has good adsorption performance for an antibody, and is particularly suitable for separating and purifying immune globulin IgG.

Description

1. Technical field [0001] The invention relates to the field of biomedical materials, in particular to the preparation of a sulfamethazine affinity membrane and its application in antibody separation and purification. 2. Background technology [0002] Antibodies are a class of proteins produced by B lymphocytes in response to antigenic stimulation. It is a globulin that can specifically bind to the corresponding antigen and produce various immune effects (physiological effects). As a special protein molecule, antibodies are often used as reagents for in vitro diagnosis, drugs for treating diseases, ligands for immunoaffinity chromatography, etc., and have a wide range of applications in the fields of life science research, biotechnology and medicine. In particular, antibodies, as the core reagents of various immunoassays, play a vital role in the sensitivity and specificity of immunoassay results. In the research of some new gene products that are closely related to the fu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01D71/82B01D71/16B01D67/00C07K1/22
Inventor 应国清朱丽易喻梅建凤王鸿陈建澍
Owner MABPLEX INT LTD
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