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

Table 2 Purification course of NFG1CG4-hFasLECD conjugated with single maleimide group containing compounds

Purification step	Sample volume (ml)		Total protein (mg)
Culture supernatant	2500		N. D.^*1
1^st Ultrafiltration plus buffer-exchange	140		3024
1^st Cation-exchange chromatography and 2^nd ultrafiltration plus buffer-exchange	7.0		69.4
After chemical modification reaction and 2^nd cation-exchange chromatography^*2	100^*3	108^*4	24.5^*3	16.5^*4

^*1Not determined. ^*2Normalized to the values starting from initial sample volume (2500 ml). ^*3Final N-Ethylmaleimide conjugated product. ^*4Final SUNBRIGHT® ME-050-MA conjugated product.

ISSN: 1472-6750