ELUTION DIAGRAM OF CYTOCHROME-C FROM ION-EXCHANGE MAGNETIC NANOPARTICLES

FERMENTATION PROFILE OF Pichia pastoris: PRODUCTION OF RECOMBINANT DROSOMYCIN

ADSORPTION ISOTHERM OF DROSOMYCIN FERMENTATION BROTH ON MAGNETIC NANOPARTICLES

SUMMARY OF DROSOMYCIN PURIFICATION USING MAGNETIC NANOPARTICLES

SDS GEL ELECTROPHORESIS USING MAGNETIC NANOPARATICLES: ELUTION PROFILES OF DROSOMYCIN

COMPARISON OF MAGNETIC NANOPARTICLES WITH OTHER PURIFICATION SCHEMES

NEW CONCEPT IN INCREASING OXYGEN TRANSFER RATE USING MAGNETIC NANOPARTICLES Magnetic particle Oleic Acid 20 nm 30-50 nm Low amount of coating Very high interfacial areas Readily recovered by magnetic filtration

SYNTHESIS OF MAGNETIC NANO-PARTICLES Aqueous solution of FeCl2 and FeCl3 NH4OH 80ºC Oleic acid coating Hitenol Fast (30 min) simple synthesis (stirred tank) Inexpensive, readily available materials

MASS TRANSFER CHARACTERIZATION IN BIOREACTORS: SULFITE OXIDATION air to mass spec Di = 10cm HL = 14.5cm [SO32-] = 0.67M [Cu2+] = 1x10-3 M VTOTAL = 20L VWORKING = 5.5L Dtank = 22cm

OXYGEN MASS TRANSFER COEFFICIENT VERSUS GASSED POWER PER UNIT VOLUME IN 20-LITER BIOREACTOR Ф: particle wt%

OXYGEN MASS TRANSFER IN E. coli FERMENTATIONS Seed culture - 100 ml LB in 500 ml shake flask - overnight culture at 37 oC, 220rpm Fermentation culture (7.5 L fermentor) - inoculation volume: 10% (v/v) - initial fermentation volume: 3 L - temperature: 37 oC - agitation speed: 600 rpm - pH = 6.8-6.9, adjusted by 4 M (NH4OH:NaOH = 2:2)

SUMMARY OF OXYGEN TRANSFER COEFFICIENT IN E. COLI FERMEMENTATION WITH AND WITHOUT MAGNETIC NANOPARTICLES Particles (w/v) Air-flow rate (L/min) Oxygen Transfer (from 7 to 10 hrs) [mmol O2/(L-h)] kLa [mmol O2/(L-h-Atm O2] Normalized kLa (air-flow rate = 2 L/min) Fermentation III none 2.0 26.68 127.0 Fermentation I 3.0 36.30 172.9 131.8 Fermentation II 2% 1.0 74.04 352.6 561.0 Fermentation IV 4% 122.84 585.0

SUMMARY AND CONCLUSIONS Micro-Bioreactors Will Began to Have Impacts in Biotechnology Processes Reducing Time in Process Development Strain Selection Medium Development Product Quality in Mammalian Cell Culture Nano-Technology Has Definite Future Product Purification Over-Coming Transport Barriers Oxygen Transfer Biocatalysis and Co-Factor Regeneration