Ultra scaledown to predict filtering centrifugation of secreted antibody fragments from fungal broth.
Extracellularly expressed anti-hen egg lysozyme single-chain antibody fragments (scFv) produced by Aspergillus awamori were recovered using filtering centrifugation. Two filtering centrifuges with 0.5- and 30-L capacities were used to represent laboratory- and pilot-scale equipment, respectively. Critical regime analysis using the computational fluid dynamics (CFD) technique provided information about the local energy dissipation rates in both units. Experimental data indicated loss of scFv activity for energy dissipation rates above about 2.0 x 10(4) W kg(-1). This loss of activity increased in the presence of gas-liquid interfaces during filtering centrifugation. An ultra scaledown filtering centrifuge with a maximum working volume of 35 mL was designed to mimic the operating conditions identified by the critical regime analysis for the laboratory- and pilot-plant-scale units. The recovered scFv activity levels and the separation performance of the three units were comparable when operated at equal maximum energy dissipation rates.
Boulding
N
N
The Advanced Centre for Biochemical Engineering, Department of Biochemical Engineering, University College London, Torrington Place, United Kingdom.
Yim
S S S
SS
Keshavarz-Moore
E
E
Ayazi Shamlou
P
P
Berry
M
M
D000818:Animals; D001230:Aspergillus / Q000235:genetics / Q000378:metabolism*; D002498:Centrifugation / Q000295:instrumentation* / Q000379:methods; D002645:Chickens; D003198:Computer Simulation; D004527:Egg Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D004867:Equipment Design; D007140:Immunoglobulin Fab Fragments; D007128:Immunoglobulin Fragments / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D007218:Industrial Microbiology / Q000295:instrumentation / Q000379:methods; D008851:Micropore Filters; D009113:Muramidase / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D010865:Pilot Projects; D011786:Quality Control; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D015203:Reproducibility of Results; D012212:Rheology; D012680:Sensitivity and Specificity; D013314:Stress, Mechanical
D004527:Egg Proteins; D007140:Immunoglobulin Fab Fragments; D007128:Immunoglobulin Fragments; D011994:Recombinant Proteins; D009113:Muramidase