A dual monoclonal antibody-based sandwich ELISA for detection of potent vaccine immunogen against Coxsackievirus B1

Fig. 1. Establishment of a DAS-ELISA method for detecting the potent immunogen against CVB1. (A) The schematic diagram of the double antibody sandwich ELISA method. (B) Screening antibody pairs that detect CVB1 mature virions. 13 × 13 antibody pairs were screened using two types of CVB1 antigens, including viral culture supernatants and heat-inactivated viral culture supernatants (56 °C for 30 min). The OD value change represents the difference in OD values between untreated viral supernatant and heat-inactivated supernatant for each antibody pair. The larger the value, the stronger the specific binding activity of the corresponding antibody pair to CVB1-M. (C) Electron microscopy images of the purified CVB1-M virons before and after heat inactivation, and the purified CVB1-E virons. D The reactivity of 5F5/8A10-HRP and 10D6/8A10-HRP antibody pairs for detecting 100 ng/mL CVB1-M, 100 ng/mL heat-inactivated CVB1-M, and 100 ng/mL CVB1-E. The values are expressed as the mean ± SD. E Binding efficiencies of antibody 5F5, 10D6 and 8A10 to CVB1-M, heat-inactivated CVB1-M or CVB1-E evaluated by direct ELISA. The values are expressed as the mean ± SD. F Samples of CVB1 stock at 3-fold serial…