Applied Microbiology: Open Access
Open Access

Abstract

Cloning and Expression of VP1 Protein for Large-Scale Production in E. coliTOP10

Elham Omer Mahgoub* and Galal M. Abdella

The VP1 capsid gene is essential for expressing a protein that enables the diagnosis of Chicken Anemia Virus (CAV) infection in chicken flocks. In this study, the prokaryotic expression vector pRSET–B expressed the VP1 gene in E. coli Top10. The VP1 protein was expressed in E. coli TOP10 as a soluble fusion protein. The solubility of the VP1 protein activates the immunogenic epitope to make it easily detected using an anti-VP1 monoclonal antibody at 50 kDa. The batch fermentation process was used to scale up the production of VP1 protein in E. coli. As a result, the recombinant VP1 protein successfully increased the yield of the expressed VP1 protein during high bacterial cell density culture. Dialysis and desalting increased the specific activity and the final yield of the VP1 protein fraction when the Tangential Flow Filtration (TFF) step was used. The statistical calculation of indirect and commercial ELISA tests indicated that indirect ELISA can compete with commercial tests with a lower amount of solution, lower costs, and higher specificity. The VP1 protein expressed in batch fermentation bacterial culture was tested as an antigen to detect antibodies to CAV in infected chickens using the developed indirect Enzyme-Linked Immunosorbent Assay (ELISA).

Published Date: 2023-09-08; Received Date: 2023-08-03