Integrated fragmentation of human IgG and purification of Fab using a reactant adsorptive membrane bioreactor separator system.
This article discusses an integrated separation-reaction-separation scheme for producing Fab fragment directly from human immunoglobulin G (hIgG) present in serum feed. The novel reactant adsorptive membrane bioreactor separator (or RAMBS) system used in the current study consisted of a stack of microporous adsorptive membranes held within a temperature controlled module. The membrane stack, in the presence of salt, selectively and reversibly adsorbed hIgG by hydrophobic interaction while allowing most other serum proteins to flow through. The bound hIgG was then fragmented by pumping a solution of papain through the reactor at controlled temperature and flow rate. The salt concentration and pH for reaction and separation were systematically optimized using pure hIgG as reactant. The Fab fragment was separated from undigested hIgG and other byproducts such as Fc fragment based on their differences in hydrophobicity. Under optimal conditions, Fab was obtained in the reaction flow through while the other proteins remained bound to the membrane, these being subsequently eluted by lowering the salt concentration. The RAMBS system in addition to being convenient from process integration and intensification points of view also showed higher catalytic efficiency of papain in comparison to that in liquid phase reactions.
Yu
Deqiang
D
Department of Chemical Engineering, McMaster University, Hamilton, Ontario, Canada.
Ghosh
Raja
R
D000327:Adsorption; D019149:Bioreactors; D006801:Humans; D006863:Hydrogen-Ion Concentration; D057927:Hydrophobic and Hydrophilic Interactions; D007140:Immunoglobulin Fab Fragments / Q000302:isolation & purification*; D007074:Immunoglobulin G / Q000378:metabolism*; D008566:Membranes; D009994:Osmolar Concentration; D010206:Papain / Q000378:metabolism; D044967:Serum / Q000737:chemistry*
D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D010206:Papain