Developing a scalable model of recombinant protein yield from Pichia pastoris: the influence of culture conditions, biomass and induction regime

Table 1 Specification of the input factors and measurable outputs for the model building experiments.

INPUT FACTORS (controlled on-line)			MEASURABLE OUTPUT (measured offline)
T (°C)	pH	DO (%)	OD₅₉₅	RFU (mL^-1)	Specific RFU (mL^-1 OD₅₉₅^-1)	Specific yield (ng mL^-1 OD₅₉₅^-1)	SD; n = 3 (ng mL^-1 OD₅₉₅^-1)
19	6	60	20.3	8651	426.2	127.9	3.2
19	8	60	0.8	1015	1268.8	380.6	3.9
19	7	30	13.1	10984	838.5	251.6	1.3
19	7	90	12.4	9259	746.7	224.0	2.1
24	6	30	24.4	8061	330.4	99.1	1.6
24	6	90	16.2	11951	737.7	221.3	5.6
24	8	30	4.7	1564	332.8	99.8	1.1
24	8	90	1.3	1954	1503.1	450.9	1.3
24	7	60	17.6	21382	1214.9	364.5	10.1
29	7	30	24.8	25392	1023.9	307.2	0.2
29	8	60	4.4	1413	321.1	96.3	1.5
29	6	60	21.7	10349	476.9	143.1	0.3
29	7	90	15.1	17495	1158.6	347.6	3.5

The input factors were temperature (T), pH and % dissolved oxygen (DO). Relative fluorescent units (RFU) and the optical density at 595 nm (OD₅₉₅) were measured in triplicate 48 h post induction. The mean values are reported for 1 mL of culture. The standard deviation (SD; n = 3) is given for the specific yield of the M24 culture, where the conversion factor from RFU to ng was determined by generating a standard curve as described in the Methods section. All experiments were performed in batch in the M24 in BMMY medium.

ISSN: 1475-2859