TABLE 2.

Optimization of human COT kinase (aa30–404) expression in Sf21 insect cells

The addition of compound 1 during fermentation improved the yield and homogeneity of COT(30–404) obtained after the first purification step (Ni-NTA yield). Fermentation volume was 0.9 l. NA, not applicable.

Compound 1	Form of addition	Time of addition	Cultivation time [h]	Ni-NTA yield	Phosphorylation in %
					0×	1×	2×
μm				mg
0	NA	NA	64	0.2–0.3	65	25	10
30	Diluted into medium, sterile filtered	At infection	63	2.4	84	16	0
10	DMSO stock diluted into culture	At infection	64	4.2	89	11	0
10	DMSO stock diluted into culture	24 h after infection	64	7	90	10	0