Application of capillary high-performance liquid chromatography to biotechnology, with reference to the analysis of recombinant DNA-derived human growth hormone.
Using capillary HPLC, femtomole amounts of recombinant DNA-derived human growth hormone (rhGH) have been successfully detected from solutions at nanomolar concentrations. The separation used capillaries of 15 cm x 320 microns I.D. and detection was with a UV absorbance detector containing a capillary Z-shaped flow-cell. A sample of rhGH that was recovered from rat serum was analyzed by capillary reversed-phase HPLC, using both acidic- and neutral-pH mobile phases, as well as by capillary ion-exchange chromatography. When compared to HPLC separations performed at flow-rates of 1 ml/min, the sensitivity of the detection was increased 200 times, without any loss in resolution. Sub-microgram amounts of rhGH were also analyzed by tryptic mapping using capillary HPLC and peptides were identified by capillary LC-MS.
Battersby
J E
JE
Genentech Inc., South San Francisco, CA 94080.
Guzzetta
A W
AW
Hancock
W S
WS
D000595:Amino Acid Sequence; D001709:Biotechnology; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004274:DNA, Recombinant / Q000378:metabolism; D013006:Growth Hormone / Q000096:biosynthesis / Q000302:isolation & purification*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D009154:Mutation; D010449:Peptide Mapping; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D013056:Spectrophotometry, Ultraviolet; D014357:Trypsin
D004274:DNA, Recombinant; D011994:Recombinant Proteins; D013006:Growth Hormone; D014357:Trypsin