Purification of human antibodies from transgenic corn using aqueous two-phase systems.
A recombinant human antibody expressed in corn was purified using aqueous two-phase extraction. The antibody was an immunoglobulin G fully unglycosylated. Using systems of different compositions and/or pHs in each of one or two partitioning stages followed by one more stage in which the antibody was precipitated at the liquid/liquid interface facilitated the removal of different impurities in each stage. The best system yields a product 72% pure (22-fold purification) with a yield of 49%. The optimum extraction was done in two partitioning stages followed by an interfacial precipitation stage using poly(ethylene)glycol/potassium phosphate systems. NaCl was added to the first stage to eliminate large molecular weight impurities. The pH in the first stage was kept at 6 but a pH of 8 was used in the second stage and in the precipitation stage.
Lee
J-W
JW
Dept. of Chemical and Biological Engineering, Missouri University of Science and Technology, Rolla, MO 65409, USA.
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D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D019075:Electrophoresis, Capillary; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000737:chemistry / Q000302:isolation & purification*; D008970:Molecular Weight; D010710:Phosphates / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000737:chemistry*; D017680:Potassium Compounds / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D012965:Sodium Chloride / Q000737:chemistry; D014867:Water / Q000737:chemistry*; D003313:Zea mays / Q000737:chemistry*
D007074:Immunoglobulin G; D010710:Phosphates; D017680:Potassium Compounds; D011994:Recombinant Proteins; D014867:Water; D011092:Polyethylene Glycols; D012965:Sodium Chloride; C013216:potassium phosphate