Hybridoma cell line secreting foot-and-mouth disease virus non-structural protein monoclonal antibody 3A10, and application thereof

Abstract

The invention discloses a hybridoma cell line secreting a foot-and-mouth disease virus non-structural protein monoclonal antibody 3A10, 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.16213, 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 thefoot-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 canbe 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 3A10 and a hybridoma cell line secreting a foot-and-mouth disease virus non-structural protein monoclonal antibody The present invention further relates to their application in distinguishing foot-and-mouth disease naturally infected animals from immunized animals, and belongs to the field of foot-and-mouth disease virus non-structural protein monoclonal antibody and its application field. 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 of wild cloven-hoofed animals. Humans ...

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 method for differential diagnosis of foot-and-mouth disease established with this non-competitive or poorly competitive monoclonal antibody is cumbersome, and more seriously, it cannot cross the barrier between animal species and cannot detect serum from any animal species in a reaction system

[0008] Due to the above-mentioned defects in the existing monoclonal antibodies used to detect foot-and-mouth disease virus to varying degrees, these monoclonal antibodies cannot establish an effective blocking ELISA technique. In addition to low sensitivity and poor broad-spectrum (that is, the conservation of epitopes), what is more serious is that a detection kit needs to be established for each animal to be detected, which lacks practical application value

Images