Expression and Purification of C-Peptide Containing Insulin Using Pichia pastoris Expression System

Table 1

Expression of recombinant human insulin in Pichia pastoris as evaluated by ELISA.

Commercial insulin		Time (hrs)	Recombinant proinsulin
Protein conc.; (µg/mL)	OD 495		Mut^+ clone		Mut clone
			OD 495	Protein conc. (µg/mL)	OD 495	Protein conc. (µg/mL)
5	0.695 0.031	0	0.447 0.015	—	0.473 0.020	—
10	0.706 0.015	24	0.523 0.010	8	0.587 0.120	11
15	0.776 0.015	72	0.749 0.023	20	0.0818 0.021	26
20	0.798 0.070	96	1.155 0.130	31	1.205 0.101	35
25	0.802 0.110	144	0.870 0.030	23	1.015 0.060	29
30	0.870 0.09

Mut: methanol utilization +ve clone.
Mut: methanol utilization slow phenotype.