Quantitative Mass Spectrometry For Biopharmaceutical Analysis

The rapid growth in biopharmaceutical development brings a myriad of new challenges for bioanalysts, not present in small molecules analysis.

The application examples below outline methods, inclusive of sample prep, separations, MS and informatics, which enable scientists to solve the most pressing and challenging analytical problems related to biopharmaceutical bioanalysis.

Waters Xevo Mass Spectrometry Solutions Integrate:

  • Chromatographic systems and mass spectrometers that combine accurate reproducible separations and maximum MS performance with intelligent, easy-to-use laboratory software converting scientific data into useful information.
  • Chemistry consumables that work with your system to maximize sensitivity, increase throughput and ensure the development of robust methods.
  • Waters Applications Support and Global Services partner with you to optimize your laboratory efficiency, help maintain system peak performance, address scientific application challenges and meet your compliance requirements to ensure the success of your laboratory and business

Download this e-book for essential information about these analyses, from general precepts to specific considerations when developing an LC–MS/MS method.

Peptide and Protein Bioanalysis Application Notebook

Peptides and Proteins are not Small Molecules; Why Treat Them the Same?

Whether you have been doing it for years or are just starting off, large molecule quantification creates a number of challenges for bioanalysts, including:

  • Sensitivity and specificity issues

  • Multiple charge states dilute signal

  • Complex fragmentation patterns

  • Endogenous proteins and peptides yield high background

  • And many, many more

Waters’ Peptide and Protein Bioanalysis Application Notebook is the most comprehensive collection of peptide and protein bioanalysis applications available. With over 48 unique analytes and with insights from 60 expert authors, you will find guidance on your most pressing bioanalytical challenges.

Application Notebook

Peptide and Protein Bioanalysis Application Notebook 

Key Challenges of Peptide and Protein Bioanalysis 

Large molecule quantitation is one of the greatest challenges that bioanalysts face. Sensitivity and specificity issues are numerous such as fewer molecules per volume, increased fragmentation and limited recovery due to complex sample prep, to name a few.

Learn more about these key challenges in this infographic.


Peptides and Proteins are Not Small Molecules: Why Treat Them The Same? 



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Xevo Family of Tandem Quadrupole Mass Spectrometers

Quantitation of Monoclonal Antibodies

Monoclonal antibodies (mAbs) comprise an expanding group of therapeutic compounds, which are used to treat cancer, auto-immune diseases, and systemic infections. Mass spectrometry is the analytical tool of choice for quantitation of mAbs, due to its selectivity, multiplexing, and dynamic range advantages over ligand binding assays (LBAs). Waters' Xevo tandem quad family, columns and chemistries have been used to conduct bioanalysis of large protein molecules, such as the monoclonal antibodies and antibody-drug conjugates (ADCs) like Bevacizumab, Infliximab, and Trastuzumab Emtansine .

ProteinWorks are a set of kits that facilitate the surrogate peptide method (protein digest), which is the most common method for mAb quantitation by LC/MS. This notebook contains key examples of accurate, sensitive, and reproducible quantitation of protein therapeutics including mAbs and ADCs.

Application Notebook


 Drug Metabolism and Pharmacokinetics (DMPK)

Due to its superior mass resolution abilities, high resolution mass spectrometry (HRMS) is now being used to overcome bioanalytical quantitation challenges facing small and large molecule development portfolios.

This includes the ability to develop validated methods in drug development, overcoming selectivity challenges and the intact analysis of large molecules


The Ever-expanding Role of HRMS in Regulated Bioanalysis: Application to Both Small Molecule and Biopharmaceutical Study Support.

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Xevo G2-XS QTof Quadrupole Time-of-Flight Mass Spectrometer

Therapeutic monitoring of Bradykinin

Bradykinin is an inflammatory mediator and an important regulator of blood pressure, kidney function and heart function, and also involved in inflammation. The ability to measure changes of this peptide hormone with high sensitivity, selectivity and accuracy, as a function of disease progression or drug treatment is highly advantageous.

LC-MS/MS using a Xevo tandem quadrupole mass spectrometer has the advantages of shorter development times, greater accuracy and precision, the ability to multiplex, and can readily distinguish between closely related analogues, metabolites or endogenous interferences.

Application Note 

Development of a Quantitative SPE LC-MS/MS Assay for Bradykinin in Human Plasma.

Microflow LC/MS for Peptide and Protein Quantitation

Microflow can facilitate sensitivity gains for a wide variety of analytes, but the greatest benefit has been for peptides and proteins.

Peptide and protein bioanalysis brings greater complexity compared to small molecule analysis. High background noise due to endogenous peptides and proteins can reduce signal and require more complex sample prep. Larger molecules have more multiply-charged species, increased fragmentation patterns, and a variety of lower abundance fragments, all of which reduce signal-to-noise ratio. Additionally, a significant number of biologics have longer half-lives than small molecules; therefore, many are dosed at low concentrations.

These issues associated with large molecule bioanalysis support the need for the sensitivity afforded by modern microflow LC/MS.


Microfluidics for macromolecules: leveraging microflow LC/MS for quantitation of therapeutic peptides

Application Note

Reducing Sample Volume and Increasing Sensitivity for the Quantification of Human Insulin and 5 Analogs in Human Plasma Using ionKey/MS

 For More Quantitative Content Visit our DMPK and Bioanalysis Knowledge Center



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