APPLICATION OF CAPILLARY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY TO BIOTECHNOLOGY, WITH REFERENCE TO THE ANALYSIS OF RECOMBINANT DNA-DERIVEDHUMAN GROWTH-HORMONE

Using capillary HPLC, femtomole amounts of recombinant DNA-derived hum an growth hormone (rhGH) have been successfully detected from solution s at nanomolar concentrations. The separation used capillaries of 15 c m x 320 mu m I.D. and detection was with a UV absorbance detector cont aining a capillary Z-shaped flow-cell. A sample of rhGH that was recov ered from rat serum was analyzed by capillary reversed-phase HPLC, usi ng both acidic- and neutral-pH mobile phases, as well as by capillary ion-exchange chromatography. When compared to HPLC separations perform ed at flow-rates of 1 ml/min, the sensitivity of the detection was inc reased 200 times, without any loss in resolution. Sub-microgram amount s of rhGH were also analyzed by tryptic mapping using capillary HPLC a nd peptides were identified by capillary LC-MS.