Hybridoma cell line secreting foot-and-mouth disease virus nonstructural protein monoclonal antibody 2h1 and its application

Abstract

The invention discloses a hybridoma cell line secreting foot-and-mouth disease virus non-structural protein monoclonal antibody 2H1 and application thereof. The present invention uses non-structural protein of foot-and-mouth disease as an antigen to immunize mice and obtains a hybridoma cell line stably secreting non-structural protein monoclonal antibody of foot-and-mouth disease virus with the preservation number of CGMCC No. 16212 through cell fusion technology screening and final screening. It has good broad-spectrum, high conservation, strong affinity, and strong competition with serum. It can be used as a detection antibody in the blocking ELISA detection method to accurately distinguish between naturally infected animals and immunized animals. The present invention further provides a blocking ELISA detection kit established by using the monoclonal antibody to detect antibodies. The serum of any animal species can be detected by using a single detection kit, which has crossed the interspecies barrier and has high reliability, It has the advantages of wide applicability, good broad spectrum, strong competition, and faster and more convenient 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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