A method for separation and purification of fmd-inactivated virus antigens using simulated fluidized bed

Abstract

The invention relates to a method for separating and purifying an FMD inactivated virus antigen by applying a simulated flow bed, wherein the method comprises the following steps: balancing a chromatographic column of the simulated flow bed with a balanced buffer solution; loading an FMD inactivated virus antigen sample into the chromatographic column of the simulated flow bed, during sample loading, simultaneously carrying out antigen sample impurity cleaning and sample desorption elution as well as chromatographic filler cleaning and regeneration and balanced liquid balancing on the chromatographic column, and continuously desorbing and eluting the collected and purified sample in the process. The simulated flow bed is applied to separation and purification of the FMD inactivated virus antigen for the first time, the continuous saturated sample loading separation operation can be realized and the sample loading amount is greatly improved; moreover, through the continuous separation and purification via multiple monomer columns, the degree of continuity is high, the separation and purification time is greatly shortened, the preparation efficiency is improved and the operation costis reduced. Moreover, the rigid filler is selected specifically, the loading capacity of the filler namely the pressure resistance is greatly improved, the use amount of the chromatographic filler isreduced, and the service life of the chromatographic column in the simulated flow bed is prolonged.

Description

technical field [0001] The invention relates to the technical field of manufacturing veterinary biological products, in particular to a method for separating and purifying FMD inactivated virus antigens using a simulated fluid bed. Background technique [0002] Foot-and-mouth disease virus (FMD) is an economically devastating viral disease transmitted among artiodactyls. In the foot-and-mouth disease virus vaccine samples, it is mainly the intact virus particles that are related to stimulating the body to produce antibodies and sensitizing lymphocytes. Its sedimentation coefficient is 146S, which is considered to have a parallel relationship with the efficiency of the foot-and-mouth disease vaccine. Internationally, the production of foot-and-mouth disease vaccines mainly adopts suspension culture technology, and the culture reaction is between 3000L-8000L. The subsequent purification process mainly uses methods including polyethylene glycol precipitation, ultrafiltration to...

AI Technical Summary

Problems solved by technology

When the above-mentioned chromatographic method is used for purification, there are often problems such as small sample loading, low packing capacity, single-line operation, long purification process time, large sample loss, large amount of waste liquid, and high treatment costs.

[0004] Simulated fluid bed technology is a purification technology developed in the 1960s. It was only applied to the separation of petrochemical products at first. In the 1990s, Seperex of France applied it to the separation and purification of pharmaceuticals and fine chemicals. For the separation and purification of sugars, isomers, chiral compounds, and biological fermentation broths, and for the separation and purification of inactivated virus antigens in suspension culture of foot-and-mouth disease, the simulated fluid bed technology has not been applied to the separation and purification of virus samples. The application of this technology has very high requirements for the fillers in the fluidized bed body, and it is necessary to find fillers that are resistant to wear, high pressure, and acid and alkali treatment. At present, the fillers of ordinary chromatography for the purification of foot-and-mouth disease inactivated virus antigens are based on dextran or The filler of agarose matrix generally has a particle size of 75um and a pressure-bearing capacity of less than 0.3MPa. Long-term acid-base treatment will easily lead to the loss of functional groups and poor purification effect, which limits the application of simulated fluidized bed technology in the inactivation of foot-and-mouth disease. Applications in the field of viral antigen purification

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