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The evolution of instrument design for protein/peptide sequence analysis has concentrated toward maximizing system efficiency and sensitivity. An integration of features from both spinning cup and solid phase type instruments together with new concepts for reactor design and reagent delivery resulted in the development of the high performance gas-liquid solid phase sequencer (Hewick et al, 1981). This instrument, made commercially available in 1981 by Applied Biosystems (ABI 470A), contained as major hardware components a novel multi-mode reactor, miniaturized reaction and conversion modules, an efficient chemical delivery system employing zero dead-volume valve blocks (Wittmann-Liebold, 1980) and microprocessor control. The addition in 1985 of an online narrow-bore gradient HPLC (ABI 120A) system for the rapid identification of PTH-amino acids (PTH-AA’s) resulted in increased sample throughput and system performance. A unique data analysis system, which in software eliminates the baseline artifact and employs a digital filter to give apparent baseline resolution for each peak in a chromatogram (Hunkapiller, 1987), has improved our ability to obtain sequence information at the low picomole level and below. |
