反相高效液相色谱对DABTH-氨基酸的分离与鉴定

Separation and Identification of DABTH-Amino Acids by Reverse-Phase High-Performance Liquid Chromatography

应用反相高效液相色谱(RP-HPLC)技术,采用等度洗脱。对20种标准DABTH-氨基酸进行了分离与鉴定,灵敏度达到pmol水平;并以溶菌酶为模式蛋白,经手工DABITC/PITC双偶联方法对其N端部分序列进行Edman降解,其降解产物DABTH-氨基酸用RP-HPLC技术进行了鉴定,蛋白质微量序列分析的灵敏度达到nmol水平.

Twenty standard DABTH-amino acids were separated and identified on a reverse-phase high-performance liquid chromatography(RP-HPLC) column with isocratic elution at a sensitivity of picomoles. By using the manual DABTH/PITC double-coupling method, we have analyzed and identified the N-terminal amino acid sequence of chicken lysozyme. This technique has been verified to be valuable for protein microsequencing at a level of nanomoles.