Table 1.

Summary of Differences in T_m (Thermal Melting Temperature) Values for Parent Protein and Site-Directed Mutants of NRRV Antigens P[8] and P[4] Produced in K. phaffii, with Respect to E. coli-Expressed Parent P[8] or P[4] Antigens, Respectively, as Measured by Differential Scanning Calorimetry (DSC) and Extrinsic Fluorescence Spectroscopy.

	NRRV Antigen	Thimerosal Addition (±)	Change in Tm (°C) (vs. E. coli Parent Protein)		Change in Tm (°C) (With vs. Without Thimerosal)
			DSC	Extrinsic Fluorescence Spectroscopy	DSC	Extrinsic Fluorescence Spectroscopy
P[8]	E. coli P[8]	–	NA		−7	−8
		+
	Pp P[8] (truncated)	–	0	−1	−8	−8
		+
	Pp P[8]	–	−1	−1	−7	−8
		+
	Pp P[8]-N85A,N151A	–	2	0	−8	−8
		+
	Pp P[8]-N85Q,N151Q,C171S	–	−5	−7	0	0
		+
P[4]	E. coli P[4]-WT	–	NA		−9	−9
		+
	Pp P[4]	–	0	0	−9	−9
		+
	Pp P[4]-C173S	–	−6	−7	0	0
		+

Samples were prepared at 0.12 mg/mL in 10 mM PBS buffer at pH 7.2. Differences in T_m values were determined both in the presence and absence of 0.01% w/v thimerosal. Each sample was measured at n = 3 and differences were determined from averaged values.

See Fig. 3 and Supplemental Table S1 for Tm ±1 SD values.

NA, not applicable.