A kind of preparation method of cytokine ELISA standard product

Abstract

The invention discloses a preparation method of standard substances of cytokine ELISA. The preparation method includes the steps of S1, precooling relative equipment for freeze drying until the temperature drops to 4 DEG C; S2, after dissolving standard protein of the cytokine in water, using a freeze-drying solution to dilute the dissolved protein until the concentration of the protein reaches the target concentration, and then obtaining a protein solution with standard concentration; S3, subpackaging the protein solution with the standard concentration into a precooled Sieling bottle, and gently putting on a cover; S4, placing the Sieling bottle containing the protein solution obtained in the step S3, in a vacuum freeze-drying machine until the protein solution is completely dried, and then obtaining the standard substances of the cytokine; the freeze-drying solution in the step S2 is prepared from, by mass, 0.5-5% of sodium caseinate, 1-10% of mannitol, 0.05-0.5% of polypyrrolidone K30 and the balance phosphate buffer with concentration of 0.01 mol / L. By using the preparation method, the standard substances of the cytokine, which have a two-year service life, can be obtained. By adopting the formula of the freeze-drying solution in the method for freeze drying, a great molding effect can be achieved, the appearance of the product is good, and undesirable phenomena such as wall hanging can not occur.

Description

technical field [0001] The invention belongs to the field of preparation of biological in vitro diagnostic reagents. Specifically relates to a preparation method of a cytokine ELISA standard product. Background technique [0002] Enzyme-linked immunosorbent assay (ELISA), as the internationally recognized gold standard for protein quantification, has the characteristics of high sensitivity and good specificity. Among them, the standard is a key to the composition of the ELISA kit, and the shelf life of the standard directly affects the shelf life of the kit. At present, the storage methods of ELISA standard products include glycerol -20°C storage method and vacuum freeze-drying method. These two methods can prolong the stability of the protein to a certain extent, but it is difficult to meet the long-term storage requirements of the kit. From practical experience, it is found that different proteins have different preservation effects on the same formula of lyophilized liq...

AI Technical Summary

Problems solved by technology

At present, the storage methods of ELISA standard products include glycerol -20°C storage method and vacuum freeze-drying method. These two methods can prolong the stability of the protein, but it is difficult to meet the long-term storage requirements of the kit.

From practical experience, it is found that different proteins have different preservation effects on the lyophilized solution of the same formula

However, the storage time of general kits on the market is half a year, which limits the use

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