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UPLC/MS/MS Method for the Quantification of Trastuzumab in Human Serum at the 5-nM Level Using Xevo TQD MS and ACQUITY UPLC H-Class System

Waters
11 Jun 2013

This application note addresses some critical challenges in the world of bioanalysis, such as developing a method to quantify protein molecules with desired sensitivity in a robust, reproducible, high-throughput LC/MS/MS system.

It demonstrates the capability of Xevo® TQD to conduct bioanalysis of large protein molecules, such as trastuzumab, a monoclonal antibody. Oasis® micro-elution plates, the ACQUITY UPLC® H-Class System, and an advanced tandem quadrupole mass spectrometer (Xevo TQD) were used for the development of a sensitive method for the quantification of trastuzumab in human serum digest.

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Waters ACQUITY UPLC - Dramatic Improvements in Speed, Sensitivity and Resolution Whether you work in methods development or quality assurance, in drug discovery and development or testing for safety in food supplies, you seek a productivity edge. With the ACQUITY UPLC® System, now you can get more information in a single, short run than you've ever seen with HPLC. Waters ACQUITY UPLC System UltraPerformance LC® (UPLC®) technology starts with unique 1.7 μm small-particle chemistries. Chromatographers no longer need to choose between speed and resolution–with UPLC you get both. Instrument Designed for Chemistry; Chemistry Designed for the Instrument The ACQUITY UPLC System has been holistically designed to match the performance needs of our innovative column chemistries with robust hardware, easy-to-use software and specialized support services. Small, pressure-tolerant particles High-pressure fluidic modules Minimized system volume Negligible carryover Reduced cycle times Fast response detectors Integrated system software and diagnostics With UPLC you have the ability to work more efficiently with higher speed, sensitivity and resolution at a much wider range of linear velocities, flow rates and backpressures to obtain superior results. The Waters ACQUITY UPLC System's high-pressure fluidics optimizes flow rates to make the most of small particle technology. The ACQUITY UPLC System's sample-handling design is designed to ensure exceptionally low carryover and reduced cycle time. And when interfaced with the Sample Organizer, it increases unattended sample capacity by ten times. High-speed detectors, both optical and mass, contribute to increased sensitivity and help manage the heightened speed and resolution requirements of UPLC. ACQUITY UPLC Systems are easily controlled, diagnosed, and monitored via a graphical system console interface. The console offers: Quick and easy access to critical instrument parameters Simple system start-up, elegant system status monitoring and predictive performance indicators to ensure maximum productivity Data management capabilities that are supported by both MassLynx™ and Empower™ software The ACQUITY UPLC System is also supported by Intelligent Device Management technology with our Connections® INSIGHT™ service, providing instrument diagnostics.

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UHPLC and HPLCHigh performance liquid chromatography (HPLC) and ultra high performance liquid chromatography (UHPLC), also known as UPLC, are analytical techniques used to separate, identify and quantitate components of complex mixtures including biological samples such as proteins and lipids as well as chemical mixtures of pesticides, drugs and oils. Both techniques are liquid chromatographic methods but differ by operating pressures (HPLC < 6000 psi < UHPLC ). Components of HPLC and UHPLC systems include columns, detectors, pumps, autosamplers and column heaters. Explore a range of UHPLC and HPLC columns for your specific sample needs including reverse phase, normal phase, ion exchange, HILIC, ion exclusion and size exclusion columns. For more specialized HPLC, explore FPLC, countercurrent LC and simulated moving bed systems. Find the best UHPLC and HPLC equipment in our peer reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Mass SpectrometryMass spectrometry (MS) is a powerful analytical technique used to identify and quantify molecules based on the mass-to-charge ratio of gas-phase ions. It provides detailed information about the structure, composition, and properties of compounds and is widely used across fields such as environmental monitoring, materials science, drug discovery and development, food and beverage testing, and wider chemical research. Key MS techniques include tandem mass spectrometry (MS/MS), liquid chromatography–mass spectrometry (LS-MS) and inductively coupled plasma (ICP-MS). Choosing from these wide range of techniques and technologies can be a daunting task, so keep up to date with scientific applications, performance expectations, and customer reviews here all in one place. Visit our product directory to receive quotes direct from the manufacturer. Cancer ResearchAlthough cancer is often referred to as a single condition, it actually consists of more than 100 different diseases. Microscopy, mass spectrometry, high throughput sequencing and flow cytometry are some of the most common techniques employed in cancer research labs.Monoclonal AntibodiesMonoclonal antibodies (mAbs) are lab-made molecules that can bind to specific antigens on cells, making them highly effective in treating cancers, autoimmune diseases, and infections. These targeted therapies are transforming the treatment landscape for many conditions. Browse our peer-reviewed product directory to find monoclonal antibodies for research and clinical applications; compare products, check reviews, and get pricing directly from manufacturers.ChromatographyChromatography is a powerful technique used to separate and analyze components of mixtures based on their chemical properties. It is widely used in fields like biochemistry, pharmaceuticals, and environmental analysis. By exploiting differences in the interaction of substances with a stationary phase and a mobile phase, chromatography enables precise purification and quantification of compounds. Whether you're working with complex biological samples or industrial chemicals, chromatography is essential for obtaining high-quality results. Browse our peer-reviewed product directory to find the best chromatography systems, compare products, read customer reviews, and get pricing directly from manufacturers.Breast CancerBreast cancer is a type of cancer that begins in the cells of the breast and can spread to other parts of the body. Early detection, accurate diagnosis, and targeted treatments are essential for improving patient outcomes. Browse our peer-reviewed product directory to find the best breast cancer research and diagnostic tools, compare products, check reviews, and get pricing directly from manufacturers.
UPLC/MS/MS Method for the Quantification of Trastuzumab in Human Serum at the 5-nM Level Using Xevo TQD MS and ACQUITY UPLC H-Class System