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| Preparation of Anti-ebola Nucleoprotein Antibodies and Establishment of Sandwich ELISA Assay |
| LIU Jun-wei1, CHANG Rui-heng1, HUI Peng1,2, DONG Shi-shang1,2, WANG Jin-feng1,2, SUN Bo1, YANG Cheng1,2 |
1. Tianjin International Joint Academy of Biomedicine, Tianjin 300457, China;
2. School of Pharmacy Nankai University, Tianjin 300071, China |
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Abstract Objective:To prepare the anti-Ebola nucleoprotein monoclonal and polyclonal antibodies, and establish a method for determining the EBOV nucleoprotein. Methods:The rabbits and mice were immunized with EBOV nucleoprotein for preparing polyclonal and monoclonal antibodies, respectively. After optimized the conditions, such as the concentration of antibodies, coating solution and so on, the sandwich ELISA was established. Results:The polyclonal antibodies and two hybridoma cell lines that could produce monoclonal antibodies were prepared. The results of Western blot showed that the binding region of both polyclonal and monoclonal antibodies were in N terminal 1~35 amino acid of the nucleoprotein. The sandwich ELISA for detecting EBOV nucleoprotein has been optimized. The linear range of detection was 31.2~1 000 ng/ml. And the limit of detection was 2.6 ng/ml. Conclusions:The high specific polyclonal and monoclonal antibodies of anti-EBOV nucleoprotein were prepared. The quantitative method to detecting EBOV nucleoprotein was established.
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Received: 23 March 2017
Published: 25 October 2017
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