Hybridoma cell line secreting foot-and-mouth disease virus non-structural protein monoclonal antibody 2H1, and application thereof

Abstract

The invention discloses a hybridoma cell line secreting a foot-and-mouth disease virus non-structural protein monoclonal antibody 2H1, and an application thereof. The hybridoma cell line stably secreting the foot-and-mouth disease virus non-structural protein monoclonal antibody, with the microbial preservation number of CGMCC NO.16212, is finally obtained through screening using a cell fusion technology by immunizing mice with the foot-and-mouth disease non-structural protein. The monoclonal antibody secreted by the hybridoma cell line has the advantages of good broad spectrum, high conservatism, strong affinity and strong competitiveness with serum, and can be used as a detection antibody in a blocking ELISA detection method to accurately distinguish naturally infected animals with the foot-and-mouth disease and immunized animals. The invention further provides a blocking ELISA detection kit established by using the monoclonal antibody as a detection antibody. The detection kit can be adopted to detect the serum of any animal species, spans the inter-species barrier, and has the advantages of high reliability, wide application range, good broad spectrum, strong competitiveness, and quickness and convenience in detection.

Description

technical field [0001] The present invention relates to a foot-and-mouth disease virus monoclonal antibody and a hybridoma cell line secreting the foot-and-mouth disease virus monoclonal antibody, and the present invention particularly relates to a foot-and-mouth disease virus non-structural protein monoclonal antibody and a hybridoma cell line secreting a foot-and-mouth disease virus non-structural protein monoclonal antibody and The application thereof, the present invention further relates to their application in distinguishing between animals naturally infected with foot-and-mouth disease and immunized animals, and belongs to the field of non-structural protein monoclonal antibodies of foot-and-mouth disease virus and their applications. Background technique [0002] Foot-and-mouth disease (FMD) is caused by foot-and-mouth disease virus (FMDV). It is a highly contagious infectious disease that mainly affects livestock such as cattle, pigs, and sheep, as well as a variety ...

AI Technical Summary

Problems solved by technology

[0005] (1) The epitope targeted by the monoclonal antibody has not been identified, and the extent of the broad spectrum of the monoclonal antibody cannot be determined: the epitope sequence and its key amino acids or important amino acids targeted by the existing monoclonal antibody are not clear, if If the key amino acids and important amino acids vary between strains, the degree of broad-spectrum will change, which cannot ensure the broad-spectrum of foot-and-mouth disease strains, and correspondingly lacks real application value;

[0006] (2) Poor broad-spectrum of monoclonal antibodies: The antigenic epitopes targeted by monoclonal antibodies are poorly conserved, resulting in no reaction with some strains of foot-and-mouth disease;

[0007] (3) Poor competitiveness of monoclonal antibodies: Some monoclonal antibodies have poor competitive ability with FMD infection serum, which makes it impossible to establish an effective blocking ELISA detection kit

The FMD differential diagnosis method established with this non-competitive or poorly competitive monoclonal antibody is not only cumbersome for warlocks, but more serious is that it cannot detect the serum of any animal species in a reaction system because it cannot cross the barrier between animal species

[0008] Because the existing monoclonal antibodies used to detect foot-and-mouth disease virus have the above-mentioned defects to varying degrees, these monoclonal antibodies cannot establish effective blocking ELISA procedures.

The ELISA detection kits established with these monoclonal antibodies, in addition to low detection sensitivity and poor broad-spectrum (that is, the conservation of epitopes), what is more serious is that it is necessary to establish an ELISA detection kit for each animal to be detected. Detection kits lack practical application value

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