An Meichen, Liu Ning
Key Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University, Harbin 150030, China.
Se Pu. 2010 Feb;28(2):180-4. doi: 10.3724/sp.j.1123.2012.00180.
A high performance liquid chromatography-matrix-assisted laser desorption/ionization time of flight/time of flight mass spectrometry (HPLC-MALDI-TOF/TOF MS) method was developed for the separation and identification of bovine lactoferricin (LfcinB). Bovine lactoferrin was hydrolyzed by pepsin and then separated by ion exchange chromatography and reversed-phase liquid chromatography (RP-LC). The antibacterial activities of the fractions from RP-LC separation were determined and the protein concentration of the fraction with the highest activity was measured, whose sequence was indentified by MALDI-TOF/TOF MS. The relative molecular mass of LfcinB was 3 124.89 and the protein concentration was 18.20 microg/mL. The method of producing LfcinB proposed in this study has fast speed, high accuracy and high resolution.
建立了一种高效液相色谱 - 基质辅助激光解吸/电离飞行时间/飞行时间质谱（HPLC - MALDI - TOF/TOF MS）方法用于牛乳铁蛋白肽（LfcinB）的分离与鉴定。牛乳铁蛋白经胃蛋白酶水解，然后通过离子交换色谱和反相液相色谱（RP - LC）进行分离。测定了RP - LC分离所得各馏分的抗菌活性，并测定了活性最高馏分的蛋白质浓度，其序列通过MALDI - TOF/TOF MS进行鉴定。LfcinB的相对分子质量为3124.89，蛋白质浓度为18.20μg/mL。本研究提出的LfcinB制备方法具有速度快、准确性高和分辨率高的特点。
