Separation and purification of antibodies in genetic engineering purification workshop
Separation and purification of antibodies in genetic engineering purification workshop
Antibody is a special protein molecule, which is used as in vitro diagnostic reagents, ligands for immunoaffinity chromatography, etc. It is widely used in life science research, biotechnology and medical fields, especially antibodies as various The core reagent of immunoassay, for immunoassay...
Text label: genetic engineering purification pharmaceutical cleanroom, antibody separation and purification
Isolation and purification of antibodies in genetic engineering purification pharmaceutical cleanroom
Antibody is a special protein molecule, which is used as in vitro diagnostic reagents, ligands for immunoaffinity chromatography, etc. It is widely used in life science research, biotechnology and medical fields, especially antibodies as various The core reagent of immunoassay plays an important role in the sensitivity and specificity of immunoassay results; no matter what method is used to generate antibodies, the antibodies need to be separated and purified in the later stage, so according to the antibodies, the appropriate separation and purification method is selected. very important.
Chromatography is the core technology of antibody separation and purification. Generally, a three-step purification strategy is adopted: crude purification, intermediate purification and fine purification. The main purpose of crude purification is to capture, concentrate and stabilize samples. Protein G affinity chromatography, and the purity of more than 95% can be achieved in one step; intermediate purification and fine purification remove specific impurities, such as host proteins, DNA, aggregates and variants, etc. Commonly used chromatography techniques include ion exchange, Hydrophobic chromatography, etc., to achieve the desired purity of the final therapeutic antibody. When purifying antibodies by gel filter, the antibody-containing sample is passed through a chromatography column filled with porous media, and the outflowing antibody fractions are collected; when the sample moves down from the top of the chromatography column, the large antibody Molecules cannot enter the gel particles and elute quickly. Small antibody molecules can enter the gel particles, and their migration in the gel column is delayed; the order of elution of antibody molecules from the gel filtration column is generally based on the size of the molecular weight from high to low. Zhongjing Global Purification can provide consulting, planning, design, construction, installation, transformation and other supporting services for genetic engineering workshops and GMP workshops.
Antibody or antibody fragment is usually extracted from natural or recombinant systems. The choice of source will affect the steps of sample processing and purification, because impurities from different sources and the required purity of target molecules are different. Usually, A column with high affinity for the target molecule will be selected, and a high-purity sample can be obtained in one step, and the contamination will be reduced as much as possible; according to different goals and conditions, a more suitable method can be used to purify the antibody to be prepared.
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