色谱
色譜
색보
CHINESE JOURNAL OF CHROMATOGRAPHY
2010年
2期
180-184
,共5页
高效液相色谱%基质辅助激光解吸电离串联飞行时间质谱%牛乳铁蛋白素%牛乳铁蛋白
高效液相色譜%基質輔助激光解吸電離串聯飛行時間質譜%牛乳鐵蛋白素%牛乳鐵蛋白
고효액상색보%기질보조격광해흡전리천련비행시간질보%우유철단백소%우유철단백
high performance liquid chromatography (HPLC)%matrix-assisted laser desorption/ionization time of flight/time of flight mass spectrometry (MALDI-TOF/TOF MS)%bovine lactoferricin%bovine lactoferrin
建立了高效液相色谱-基质辅助激光解吸电离串联飞行时间质谱(MALDI-TOF/TOF MS)分离鉴定牛乳铁蛋白素(bovine lactoferricin,LfcinB)的方法.采用胃蛋白酶酶解牛乳铁蛋白,酶解液离心后取上清液,经过离子交换色谱、反相液相色谱、透析等技术分离,对分离得到的目标产物进行抗菌活性分析、蛋白含量测定和MALDI-TOF/TOF MS鉴定.分离得到高活性的LfcinB,其相对分子质量为 3 124.89,蛋白含量为18.20 μg/mL.本方法具有精度高、分析速度快和分辨能力强等优点,是其他传统的分析鉴定LfcinB方法所无法比拟的,为进一步研究LfcinB奠定了基础.
建立瞭高效液相色譜-基質輔助激光解吸電離串聯飛行時間質譜(MALDI-TOF/TOF MS)分離鑒定牛乳鐵蛋白素(bovine lactoferricin,LfcinB)的方法.採用胃蛋白酶酶解牛乳鐵蛋白,酶解液離心後取上清液,經過離子交換色譜、反相液相色譜、透析等技術分離,對分離得到的目標產物進行抗菌活性分析、蛋白含量測定和MALDI-TOF/TOF MS鑒定.分離得到高活性的LfcinB,其相對分子質量為 3 124.89,蛋白含量為18.20 μg/mL.本方法具有精度高、分析速度快和分辨能力彊等優點,是其他傳統的分析鑒定LfcinB方法所無法比擬的,為進一步研究LfcinB奠定瞭基礎.
건립료고효액상색보-기질보조격광해흡전리천련비행시간질보(MALDI-TOF/TOF MS)분리감정우유철단백소(bovine lactoferricin,LfcinB)적방법.채용위단백매매해우유철단백,매해액리심후취상청액,경과리자교환색보、반상액상색보、투석등기술분리,대분리득도적목표산물진행항균활성분석、단백함량측정화MALDI-TOF/TOF MS감정.분리득도고활성적LfcinB,기상대분자질량위 3 124.89,단백함량위18.20 μg/mL.본방법구유정도고、분석속도쾌화분변능력강등우점,시기타전통적분석감정LfcinB방법소무법비의적,위진일보연구LfcinB전정료기출.
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 μg/mL.The method of producing LfcinB proposed in this study has fast speed,high accuracy and high resolution.
