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Figure 7 | BMC Biotechnology

Figure 7

From: 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 modifications

Figure 7

Purification of NFG1CG4-hFasLECD conjugated with N -Ethylmaleimide/SUNBRIGHT ® ME-050MA. a) Elution profiles in cation-exchange chromatography. Used column: Resource S 1 ml. Panels: left, before reaction (non-reduced); center, after reaction with N-Ethylmaleimide and quenching; right, after reaction with SUNBRIGHT® ME-050MA and quenching. The fractions shown in underbars were collected as the final products. NaCl concentrations under each principal peak eluting condition are described. b) Elution profiles of fractionated products in size-exclusion chromatography. Used column: Superdex 200 10/300 GL. Elution buffer: 50 mM sodium acetate plus 150 mM NaCl (pH 5.6). Flow rate: 0.5 ml/min. Panels: left, molecular-weight standards mixture (A, Aldolase; B, Ovalbumin; C, Ribonuclease A); center, N-Ethylmaleimide modified product; right, SUNBRIGHT® ME-050MA modified product. Retention times of each peak are described. c) SDS-PAGE analysis of fractionated products. Lanes: M, molecular-weight size-markers; a, tag-free hFasLECD (0.15 μg); b, N-Ethylmaleimide adduct of NFG1CG4-hFasLECD (0.15 μg); c, SUNBRIGHT® ME-050MA adduct of NFG1CG4-hFasLECD (0.15 μg).

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