Table 3. Mutational stability prediction of substituted amino acids at B-cell epitopes of E.

Mutations	mCSM	Effect	SDM	Effect	DUET	Effect	I-Mutant2.0	Effect
T309A	- 0.464 Kcal/mol	Destabilizing	0.12 Kcal/mol	Stabilizing	-0.22 Kcal/mol	Destabilizing	-0.15 Kcal/mol	Decreased stability
T335A	- 0.596 Kcal/mol	Destabilizing	0.12 Kcal/mol	Stabilizing	-0.337 Kcal/mol	Destabilizing	0.33 Kcal/mol	Increased stability
R335T	- 0.304 Kcal/mol	Destabilizing	-0.38 Kcal/mol	Destabilizing	-0.173 Kcal/mol	Destabilizing	NA	NA
V391I	- 0.375 Kcal/mol	Destabilizing	0.08	Stabilizing	-0.022	Destabilizing	-0.25 Kcal/mol	Decreased stability
D393E	- 0.38 Kcal/mol	Destabilizing	0.06 Kcal/mol	Stabilizing	-0.149 Kcal/mol	Destabilizing	NA	NA