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119 results about "DIAGNOSTIC ANTIGENS" patented technology

Foot-and-mouth disease virus 2C3ABC recombinant protein as well as preparation method and application thereof

InactiveCN104788547AShow detection specificityShorten the emergency response time for prevention and controlSsRNA viruses positive-senseVirus peptidesBacteroidesSolubility
The invention discloses a foot-and-mouth disease virus (FMDV) 2C3ABC recombinant protein as well as a preparation method and application thereof, and belongs to the field of pharmaceutical biotechnology. According to the invention, the mutation of the following sites is performed on the basis of an original FMDV 3C protease gene: Cys142-Ser, Cys163-Gly. FMDV recombinant protein 2C3ABC is expressed as an inclusion body in the bacterial cytoplasm, and is subjected to separation, denaturation, renaturation and multi-step purification, to obtain a complete and enzymolysis-free FMDV nonstructural protein mu2C3ABC, wherein the protein has solubility and complete antigenicity, and has a molecular weight of 72 kDa. The protein, as a diagnostic antigen, is prepared into chromatographic strips, has sensitivities of 98.4% and 100% respectively in the detection of FMDV experimentally infected pigs and naturally infection-free pigs, and has specificities of 100% and 98% respectively in the detection of naturally infection-free pigs and vaccine-immunized pigs. Compared with commercial kits Ceditest and UBI, the FMDV 2C3ABC recombinant protein has a high degree of consistency, can be used to distinguish infected animals and immunized animals, wherein K = 0.823 (p is smaller than 0.05).
Owner:吕宏亮 +2

Method for purifying recombinant VP1 antigen of enterovirus type 71 viruses

The invention relates to a method for purifying a recombinant protein of the gene engineering and aims to provide a method for purifying a recombinant VP1 antigen of enterovirus type 71 viruses. The method comprises the following steps of: expressing and producing a recombinant VP1 protein of the enterovirus type 71 viruses by Escherichia coli obtain an inclusion body of the protein; dissolving the inclusion body in solubilizing liquid; and dropwise adding the solution of the inclusion body into renaturation solution under the conditions of water bath and magnetic stirring and then carrying out stirring renaturation on the mixture for 48 hours. The invention discloses the method, which can make the protein existing in a form of the inclusion body become a soluble protein by protein renaturation so as to greatly improve recovery rate of an expression product. The method adopts affinity chromatography for purification in one step. The purity is over 95 percent. The method for purifying the recombinant VP1 antigen of the enterovirus type 71 viruses has simple, convenient and time-saving operation, is favorable for large-scale industrial production, has the advantages of low cost, simple and convenient process, high purity of the purified protein and the like and can provide the diagnostic antigen for development of an EV type 71 diagnostic reagent and seroepidemiological survey.
Owner:HANGZHOU ZHEDA ZIJIN BIOTECH
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