PREPARATIVE CHROMATOGRAPHY OF PROTEINS

HPLC HAS BEEN SHOWN TO BE AN EXCELLENT METHOD FOR THE ANALYSIS OF PROTEINS AND PEPTIDES SUCH AS VARIANT HEMOGLOBINS, INTERFERON, AND CREATINE PHOSPHOKINASE; HOWEVER, THERE ARE NO SIMILAR SUPPORTS AVAILABLE FOR USE ON A PREPARATIVE SCALE. THIS APPLICATION DESCRIBES THE SYNTHESIS OF A SERIES OF BONDED PHASE LIQUID CHROMATOGRAPHY SUPPORTS FOR THE PREPARATIVE PURIFICATION OF PROTEINS AND PEPTIDES. SUCH SUPPORTS WOULD HAVE HIGH RESOLUTION AND HIGH CAPACITY, YET ALSO BE STABLE AND RIGID. THE SPECIFIC AIM OF PHASE I WOULD BE TO FIND THE OPTIMUM SUPPORT, IN TERMS OF RESOLUTION AND CAPACITY, FOR PREPARATIVE CHROMATOGRAPHY OF PROTEINS BY BONDING A POLYMERIC POLYAMINE COATING, A WEAK ANION-EXCHANGER, TO MANY SAMPLES OF POROUS SILICA OR OTHER RIGID SUPPORTS. PROTEIN SAMPLES SUCH AS OVALBUMIN, SERUM ALBUMIN, AND SERUM WOULD BE ANALYZED ON THE COLUMN TO TEST FOR CAPACITY AND RESOLUTION. IN PHASE II OF THE RESEARCH, THE SYNTHESIS OF CATION-EXCHANGE, HYDROPHOBIC, AND ANION-EXCHANGE BONDED PHASES WOULD BE OPTIMIZED ON THE SUPPORT CHOSEN IN PHASE I TO PROVIDE THE HIGHEST POSSIBLE LOADING CAPACITY FOR PROTEINS. THE GREATEST COMMERCIAL USE FOR SUCH PRODUCTS WOULD BE BY BIOTECHNOLOGICAL OR PHARMACEUTICAL COMPANIES WHO ARE TRYING TO PURIFY PROTEINS AND PEPTIDES ON A LARGE SCALE.