Miniaturized protein microsequencer with PTH amino acid identification by capillary electrophoresis II. A syringe-pump-based system for covalent sequencing

摘要

A miniaturized protein and peptide microsequencer consisting of a fused silica capillary reaction chamber is described. Extremely small volumes of reagents, 2 μl or less, were delivered directly to the reaction chamber through individual fused silicacapillary lines using low pressure syringe pumps. Other than an argon gas controller, no valves were used in the delivery system. The short flow path and very low dead volume were achieved by directly connecting the narrow-bore capillaries to the reaction chamber. This configuration minimized side reactions. The elimination of valves, as well as the use of capillaries for the reaction chamber and delivery lines, greatly simplified the construction of the sequencer. The performance of the sequencer wasevaluated by sequencing 8-33 picomoles of myoglobin and insulin chain B that were covalently attached to Sequelon-DITC and Sequeion-AA membranes.