Molecular dynamics simulation of protein secondary structural and dynamic properties showed the tag-less CFA/I/II/IVMEFA proteins maintained stable secondary structure during the entire simulation, indicated by peptide segments maintained same structure (the same color) as simulation time evolved (Figure 3). The root mean square deviation (RMSD) from the initial model gradually increased to 0.29 nm but became stabilized after 70 ns of simulations, indicating that the simulation reached the equilibrium. Variable root mean square fluctuation (RMSF) calculated to quantify conformational flexibility indicated that all seven epitope domains of the tag-less MEFA protein were stable during the simulation. Little dynamics was observed from the target epitope regions (Figure 4). This suggested that insertion of these epitopes did not appear to alter the stability of the overall structure of the backbone.