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詳細(xì)介紹
單克隆抗體與重組蛋白質(zhì)越來越多地被作為生物治療性制劑。相應(yīng)地，LC和LC/MS方法也作為有力的分離技術(shù)而占據(jù)了重要地位，用于支持蛋白鑒定和穩(wěn)定性測(cè)試。沃特世蛋白分離技術(shù)(PrST：Protein Separation Technology)產(chǎn)品包括反相、離子交換和分子篩色譜柱，能夠很好的應(yīng)對(duì)這些分離挑戰(zhàn)。此外，我們基于HPLC的BioSuite、SEC、IEX、HIC和反相HPLC柱為這些大分子分離提供了杰出的色譜性能。
ACQUITY UPLC SEC系統(tǒng)解決方案基于的亞乙基橋雜化(BEH)
二醇包膜顆粒柱技術(shù),能將蛋白質(zhì)或肽單體與其聚合物快速分離并進(jìn)行準(zhǔn)確定量，這是生物制藥領(lǐng)域中各大客戶購買ACQUITYUPLC SEC系統(tǒng)解決方案的關(guān)鍵原因所在。
■與使用軟凝膠或>5_m硬顆粒的傳統(tǒng)SEC方法相比，該技術(shù)可將聚合物分析的準(zhǔn)確測(cè)定速度提高10倍之多，且所消耗的洗脫液更少。
■顯著降低了對(duì)高鹽濃度流動(dòng)相的需求，減少了蛋白質(zhì)/肽和SEC顆粒表面之間的離子交換次級(jí)作用。
■用相關(guān)蛋白質(zhì)和肽對(duì)填料批次進(jìn)行質(zhì)控測(cè)試，有助于確保達(dá)到的批間穩(wěn)定性，從而提供了驗(yàn)證方法的可靠程度。
■擁有三種孔徑選擇：125?、200?和450?，適用于不同分子量級(jí)別的生物藥項(xiàng)目。
2010年，沃特世推出了1.7µm的ACQUITY UPLC Protein BEH SEC色譜柱，用于在沃特世低擴(kuò)散的UPLC儀器上獲得性能。與傳統(tǒng)的5-8 µm SEC色譜柱相比，ACQUITYUPLC Protein BEH SEC色譜柱在更短時(shí)間內(nèi)表現(xiàn)出了的組分分離度。2015年開始，沃特世相繼推出化學(xué)性質(zhì)一致的XBridge Protein BEH SEC125?、200?和450?, 3.5, 2.5 µm色譜柱，可在常規(guī)HPLC平臺(tái)上提供相當(dāng)?shù)慕M分分離度。
Analysis of Biomolecules by Size- Exclusion UltraPerformanc e Liquid Chromatography
In the production of biopharmaceuticals, there may be different analytical requirements for groups performing clone section, formulations and stability, and quality control (QC). Depending on the goal of the separation, methods may be optimized for fast analysis time, highest possible resolution, and/or reproducibility. Size-exclusion (SEC) chromatography is often used throughout the biopharmaceutical production process for the analysis of proteins and their aggregates. While SEC has traditionally been used in conjunction with low pressure HPLC instrumentation, the advent of UPLC® Technology and new sub-2 µm packing materials allows for substantial improvements in chromatographic resolution and throughput. This application note will demonstrate the use of the new ACQUITY UPLC® SEC solution for the improved detection and/ or faster analysis of protein aggregates in biopharmaceuticals.
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