Figure 8From: Improved production of recombinant human Fas ligand extracellular domain in Pichia pastoris: yield enhancement using disposable culture-bag and its application to site-specific chemical modificationsChemical modification of NFG1CG4-hFasLECD with SUNBRIGHT ® DE-100MA. a) Chemical structure of SUNBRIGHT® DE-100MA. b) Fractionation of the reaction mixture using size-exclusion chromatography. Used column: Superdex 200 10/300 GL. Elution buffer: 50 mM sodium acetate plus 150 mM NaCl (pH 5.7). Flow rate: 0.5 ml/min. Panels: left, elution profile; right, SDS-PAGE analysis of each peak fraction. Retention times of each peak are described (left panel). The fractions shown in underbars were collected as the final products. Lanes: a, peak I; b, peak II; c, peak III; d, peak IV (right panel). The protein bands composed only of the NFG1CG4-hFasLECD subunits conjugated with SUNBRIGHT® DE-100MA are arrowed. c) Characterization of the fractionated products. Panels: left and center, elution profiles of peak II (left) and peak III (center). Retention times of each peak are described. Right panel, SDS-PAGE analysis. Lanes: M, molecular-weight size-markers; a, peak II fraction (0.3 μg); b, peak III fraction (0.3 μg).Back to article page