Heterosandwich Immunoassay for the Detection of Ebola Viral Antigen VP40

摘要

Background: Ebola is one of the major public health concern with mortality rates greater than 90% in infected individuals. Till 29 November 2015, 23,687 cases of Ebola disease were seen worldwide, among which 11,315 died. There is a definite and urgent need to develop a rapid, reliable, inexpensive and sensitive diagnostic to help manage the disease from initial stages. Our research aims to develop next generation bispecific antibody based assay for the diagnosis of the Ebola virus VP40 structural protein to be used at point of care and clinical settings. Methods: Recombinant VP40 was used to immunize chicken and mice for the development of highly specific IgY antibodies and MAbs. Second generation bispecific antibodies were developed by fusing MAbs with dual specificities. Standardized combinations of IgY, mAbs and bsmAb has been used to design a sensitive Ebola detection assay. Results: Successfully standardized immunoassay with novel ebola antibodies with significant improvement in detection limit over existing assays. The assay has less steps without compromising efficiency and can be used by minimally trained personnel. Conclusion: The uniqueness of our study is the use of inexpensive chicken IgY antibody in combination with high affinity bispecific antibodies, monoclonal antibodies to increase the sensitivity of the detection system for Ebola infectious disease. The diagnostic assay developed has precision, specificity and able to detect antigens in the nanogram levels. The Ebola VP40 immunoassay could be an efficient method of screening suspected individuals during another outbreak.