一种蛋白质可以消化成数百条肽片段，仪器的分辨率是您分析结果的信心所在。与传统的高效液相色谱（HPLC）方法相比，该项UPLC^®肽分析解决方案可在较短时间内提供优秀的分辨率和灵敏度。

沃特世公司的专业人员考虑了肽分析诸多方面的因素，包括从化学、仪器控制、软件、可用性方法、文件资料、应用技术支持和远程维护等，创建了UPLC肽分析解决方案。我们的端到端系统将独特的肽分离技术色谱柱与先进的UPLC/MS性能结合到一起。其结果是复杂的蛋白质消化后较清晰的分辨率，尤其是糖肽清晰度以及较高的通量。

UPLC肽分析解决方案：

• UNIFI生物制药平台解决方案
• ACQUITY UPLC系统和ACQUITY UPLC H-Class Bio系统
• ACQUITY UPLC光二极管阵列检测器（PDA）和ACQUITY QDa质谱检测器
• 仪器控制软件：MassLynx、Empower及UNIFI
• ACQUITY UPLC BEH130和BEH300多肽分离技术色谱柱，130Å或300Å孔径，且优化用于三氟乙酸（TFA）或甲酸（FA）改性的流动相
• MassPREP肽标准品
• 高灵敏度，可获得精确UPLC/MS数据的Xevo G2-XS QTof质谱仪
• BiopharmaLynx™应用管理软件，用于进行自动数据分析和高分辨率肽质谱（MS）分析数据的报告
• ProteinWorks蛋白质定量分析试剂盒

UPLC肽分析解决方案是沃特世公司的一套成熟的性能解决方案（APS），是能够将工业先进技术-仪器控制、化学、软件、资质工具和对某种特殊的应用型性能进行具体优化的服务等系统的一员。该解决方案由系统科学家为应用科学家设计，减少了您在应用中选择解决方案时的推测工作。

肽分析成功的关键

肽图分析是一项用于全面表征生物制药产品的重要技术。在肽图中，每个肽碎片必须能够分离成单个谱峰，而蛋白质酶解物固有的复杂性为这项色谱分析工作带来了巨大的挑战。

除了蛋白质酶消化过程中产生的大量肽碎片之外，其它肽结构（翻译后修饰、氧化等）的数量也十分庞大，分析起来同样棘手。实现足够的肽图分离度是一个十分漫长的过程，效果良好的方法通常运行时间较长，并且可能无法达到预期的稳定性。现在，沃特世的UPLC^®肽分析解决方案能够出色地解决所有这些问题。

借助这套专业系统配置，您将体验到：

• 更出色的分离度、重现性和灵敏度
• 各种肽变异体的定性和定量分析均可实现色谱分离，并且在极端摩尔比下峰形也不会失真
• 动态范围十分宽泛，确保即使在低定量限条件下也能准确检出具有极端摩尔比的肽
• 灵活的肽图分析，准确表征蛋白质的关键因素
• 方法开发具有多种选择，所使用的色谱柱均经过严格的质量控制检测和验证，通过肽酶解物确保分析重现性

Significantly Higher Resolution

UPLC provides notable improvements in peptide mapping when compared to HPLC. Exceptional resolution is possible because separations obtained through the use of small particles result in narrower peaks. Just as significant, the surface chemistry of our Peptide Separation Technology packing material provides exceptional peak shape for a wide range of peptide structures.

This remarkable column performance can only be achieved with the optimized instrumentation of the ACQUITY UPLC System, which features reduced system volumes, minimal detector band-broadening and accelerated data acquisition rates required to preserve high-efficiency separations, while maximizing sensitivity.

Comparison of HPLC vs. UPLC resolving power

UPLC can provide twice the resolving power, as shown in the comparison of the separation of a tryptic digest of yeast alcohol dehydogenase by HPLC and UPLC.

Qualitative and Quantitative Analysis of Variant Peptides

• All the elements of the UPLC Peptide Analysis Solution contribute to the successful qualitative detection and quantitative measurement of trace peptides:
• The performance of ACQUITY UPLC based fluidics ensures retention time reproducibility
• High-resolution separations of both large and small peaks are essential for accurate integration
• Peptide Separation Technology particles deliver chromatographic resolution and undistorted peak shape at extreme molar ratios
• The system’s advanced optical and mass detection technologies have a wide dynamic range ensuring that peptides, at these extreme molar ratios, are detected even at the lowest limits of quantitation

Deamidation of the peptide containing Asn and Gly, T-16, shifts the retention time and resolves into three unique chromatographic peaks

A tryptic digest of bovine hemoglobin was spiked with the peptide mixture as a surrogate to represent modified peptides and separated with a Peptide Separation Technology Column. The surrogate peptide concentrations were 0.2 to 2% of the digest concentration on a molar basis. The top panel shows the peptide map of bovine hemoglobin with peptide standards spiked at 2% molar basis. A total injection on-column of 200 pmol of hemoglobin digest was made. The bottom panel focuses on the elution position of the surrogate. Thus a small trace peptide can be identified and quantitated, as shown, down to a 0.2% level. To achieve this sensitivity resolution, peak shape must be maintained over a wide mass load.