Optimization of production of the anti-keratin 8 single-chain Fv TS1-218 in Pichia pastoris using design of experiments

Table 1 Box-Benhken design matrix and experimental results for the different factors and their effect on the responses.

Experiment	pH	Temp°C	MeOH %	ABS	ΔWCW (%)	Viability %	pH_final
N1	6	10	1.25	0.66	+4,1	99.73	5.2
N2	8	10	1.25	1.05	+2,2	99.85	6.5
N3	6	20	1.25	0.10	-5,3	99.02	5.1
N4	8	20	1.25	0.02	-1,0	99.34	6.7
N5	6	15	0.5	0.45	-5,2	99.56	4.5
N6	8	15	0.5	0.98	-1,5	99.37	6.6
N7	6	15	2.0	0.75	-4,4	99.62	5.0
N8	8	15	2.0	0.67	-1,8	99.56	6.6
N9	7	10	0.5	1.13	+3,5	99.45	6.2
N10	7	20	0.5	0.22	-5,0	99.02	6.2
N11	7	10	2.0	1.07	+2,2	99.67	6.3
N12	7	20	2.0	0.13	-5,2	99.16	6.4
N13*	7	15	1.25	1.16	-1,3	99.68	6.3
N14*	7	15	1.25	1.18	-1,1	99.74	6.2
N15*	7	15	1.25	1.12	-0,8	99.63	6.2
N16*	7	15	1.25	1.27	+1,2	99.66	6.2

*Centerpoint experiments
Temp: temperature (°C); MeOH: methanol concentration (%, v/v); ABS: absorbance at 405 nm with subtracted background (650 nm) reflecting TS1-218 yield; ΔWCW: change in wet cell weight (%) after 72 h of induction compared to WCW at start of induction (initial mean WCW: 564.4 ± SD 0.006 mg); pH_final: pH of the culture medium after 72 h of induction.

ISSN: 1475-2859