One-step affinity purification of fusion proteins with optimal monodispersity and biological activity: application to aggregation-prone HPV E6 proteins

Table 2 Summary table for purification results: Monomer are defined based on their elution volume, as it follows: MBP-16E6mut: V_e = 1.92 ± 0.01 mL; MBP-8E6: V_e = 1.99 ± 0.04 mL Oligomers are defined as all protein species eluted between void volume (V₀) (included) and monomer elution volume (excluded)

	Monomer (%)	Oligomer (%)	Total A280 GF peak area (mAU × mL)
A: MBP-16E6mut
(1) Filter elution
N1	95	5	256
N2	87	13	243
N3	52	48	167
N4	85	15	480
N5	30	70	116
N6	68	32	345
A	10	90	263
(2) Pipetting elution
N1	86	14	366
N2	88	12	125
N3	60	40	125
N4	72	28	128
N5	39	61	39
N6	69	31	155
B: MBP-8E6
(1) Filter elution
N1	19	81	467
N2	30	70	113
N3	12	88	230
N4	22	78	256
N5	7	93	194
N6	9	91	404
A	16	84	486
(2) Pipetting elution
N1	71	29	301
N2	56	44	117
N3	28	72	227
N4	56	44	121
N5	16	84	156
N6	74	26	89
(3) Optimization of lysate incubation time
N1
5 min	74	26	185
30 min	77	23	160
2 h	56	44	220
N2
5 min	92	8	25
30 min	71	29	38
2 h	77	23	138

Quantification of monomeric and oligomeric species is based on the integration of elution peak (absorbance at 280 nm), with estimated uncertainty of ± 15% for monomer/oligomer ratio and ± 57 mAU × mL for total A80 peak area based on duplicated experiments

ISSN: 1475-2859