Hybrid tumor cell strain capable of secreting sugarcane-mosaic-virus-resistant monoclonal antibody and application of monoclonal antibody of hybrid tumor cell strain

Abstract

The invention discloses a hybrid tumor cell strain capable of secreting a sugarcane-mosaic-virus-resistant Yunnan strain monoclonal antibody and application of the monoclonal antibody of the hybrid tumor cell strain. Purified sugarcane mosaic virus (SCMV) particles serve as antigens to conduct immunity on BALB / c mice, cell fusion, screening and cloning are conducted, a hybrid tumor cell strain 22A6 which can conduct passage stably and secrete the SCMV Yunnan strain monoclonal antibody is obtained, and the preservation number is CGMCC No. 12007. The indirect ELISA titer of ascites of the monoclonal antibody secreted by the hybrid tumor cell strain reaches 10<-7>, and the antibody type and subclass are an IgG1 light chain and a kappa light chain. The antibody has a specific immunity response with SCMV shell protein of 36kDa; the 22a6 monoclonal antibody is utilized as the core to establish an ACP-ELISA method and a dot-ELISA method for detecting SCMV in corn, and the sensitivity of the two methods for detecting diseased leaves reaches 1: 163840-time attenuation (w / v, g / mL) and 1: 5120-time attenuation (w / v, g / mL). Obtaining of the hybrid tumor cell strain 22A6 and the monoclonal antibody thereof and establishment of a serological detecting method provide material and technological support for diagnosis and detection of the viral disease, breeding for disease resistance and scientific prevention and control.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a hybridoma cell line secreting anti-sugarcane mosaic virus monoclonal antibody and the application of the monoclonal antibody. Background technique [0002] Sugarcane mosaic virus (SCMV) is the pathogen that causes corn dwarf mosaic and sugarcane mosaic. Corn dwarf mosaic disease first occurred in Ohio State, USA in 1963, and then as many as 37 states in the United States have reported the virus disease one after another. In 1968, maize dwarf mosaic disease broke out in Xinxiang and Anyang, Henan, my country, causing huge losses to the local area. The grain loss in Hui County alone reached 33.53% of the total grain output of the county in that year. The disease, such as Heilongjiang, Liaoning, Beijing, Shanghai, Zhejiang, Guangxi and Taiwan provinces (municipalities), among which the harm is more serious in North China and Northwest China. Sugarcane mosaic disease is a worldwide su...

AI Technical Summary

Problems solved by technology

The current early warning and monitoring system for sugarcane mosaic virus has not yet been established, and even many major endemic areas have not carried out virus prediction, early warning, prevention and control work

In addition, currently only field symptom observation, RT-PCR detection, electron microscope observation and other methods are used to detect the virus, but these detection methods are inefficient and not suitable for large-scale sample detection and field application

The serological method is simple to operate, has good specificity, and can be used for large-scale detection, but the method must rely on specific and sensitive virus antibodies

Images