Generation And Characterization Of Specific Monoclonal Antibodies Against Monkeypox Virus

Orthopoxviruses are a genus of DNA viruses of which four species are known to cause disease in humans: vaccinia virus(VACV), cowpox virus (CPXV), variola virus (VARV), and monkeypox virus (MPXV). Monkeypox virus is a zoonotic virus,endemic to central and Western Africa that can cause smallpox-like symptoms in humans. The virus was initially identified in non-human primate rash lesions in 1958 and first identified in humans in 1970 during the smallpox eradication efforts. To date,4437 cases have been reported, mainly concentrated in the West African tropical rain forest area and the mortality rate was 4% to 10%. In the absence of smallpox, human infections with monkeypox virus constitute the most significant communicable Orthopoxvirus-associated illnesses extant in the world today. Both variola and monkeypox viruses are considered threat agents that could be used unlawfully for acts of bioterrorism. With monkeypox, there is the added question of its persistence in nature, and its potential for expansion in a world that is now more than 30 years without smallpox. A significant expansion of monkeypox in the world today could pose many of the same challenges as a resurgence of smallpox.Zoonotic transmission and human to human transmission indicated that it's necessary to establish a specificity ,sensitivity and convenient diagnostic method for MPXV. Current diagnostic tests including virus-specific PCR, which requires sensitive reagents and equipment. Chinese Center for Disease Control and Prevention shoulders the responsibility of preventing human from monkeypox virus infection.The study aims to establish monkeypox vius specific immunological detection method in China CDC.The double stranded DNA monkey pox virus consists of 196kb genome .The genome is divided into A ?P 16 segments and has high homology with other orthopoxvirus genomes. The A29 protein encoded by the A29 gene in region A is the envelope protein of the virus, which mediates the recognition of the virus against the host cell. Peptide A2917?49 on the A29 protein has an epitope of monoclonal antibody mAb 69-126-3-7.Based on the above research results : for establishing monkey poxvirus-specific immunofluorescence detection method,synthetic monkeypox virus A2917?49 polypeptide was used to immune BALB/c mice. Monkeypox virus monoclonal antibodies were generated through fusion, cloning and screening techniques..The main results are as follows:Monkeypox virus A29 protein, vaccinia virus A27 protein and vaccinia virus 162 protein were successfully expressed. The purity were 90.6%, 93.8% and 96.1%..And 8 strains monkeypox virus specific monoclonal antibodies were screened by the three purified proteins. Immunofluorescence assay showed that the VACV and EVCV had no cross reaction with monoclonal antibody.