Bläsi U, Linke R P, Lubitz W
Institute of Genetics and Microbiology, University of Munich, F.R.G.
J Immunol Methods. 1988 Apr 6;108(1-2):209-12. doi: 10.1016/0022-1759(88)90421-8.
A simple, rapid and reproducible method is presented for direct determination of the substitution ratio of a carrier protein with a synthetic nonradioactively labeled peptide. The peptide was covalently linked by a thiol group of a cysteine residue to the immunogenic carrier protein using the heterobifunctional reagent N-succinimidyl 3-(2-pyridyldithio)propionate. The substitution ratio was determined after reductive cleavage of the intermolecular disulfide bond between peptide and carrier and the amount of carrier and peptide quantitated directly by calibrated HPLC analysis within 15 min.
本文介绍了一种简单、快速且可重复的方法，用于直接测定载体蛋白与合成的非放射性标记肽的取代率。使用异双功能试剂N-琥珀酰亚胺基-3-(2-吡啶二硫代)丙酸酯，通过半胱氨酸残基的巯基将肽共价连接到免疫原性载体蛋白上。在还原裂解肽与载体之间的分子间二硫键后，测定取代率，并通过校准的高效液相色谱分析在15分钟内直接定量载体和肽的量。
