Large Molecule Quantification

Address the quantification of peptide- and protein-based biotherapeutics and biomarkers.

Peptide Optimization Using Skyline and the Xevo TQ-XS

This technology brief demonstrates the workflow for peptide MRM optimization using the Xevo TQ-XS Triple Quadrupole Mass Spectrometer, Skyline software and phosphorylation B peptide standard.

Why Should Bioanalysts Use Microflow LC-MS?

Why are scientists at leading companies like GlaxoSmithKline, Merck, and Pfizer using microflow LC-MS for more routine applications? The answer is simple: an overarching need to do more with less. They are working with more complex analytes, with limited sample, often requiring both improved sensitivity and high throughput. Although microflow LC-MS has been perceived as a tradeoff between high sensitivity and throughput, recent advances are yielding flexible systems that enable analysts to collect highly sensitive data with cycle times comparable to analytical scale.

Peptide & Protein Bioanalysis Application Notebook (2017 Edition)

This comprehensive application notebook addresses a variety of biologic bioanalysis challenges, with complete quantitative solutions. Download the notebook and see why Waters is leading the industry in biologic bioanalysis.

Sensitive and Reproducible LC-MS Quantification of C-Reactive Protein in Plasma: A Potential Biomarker of Inflammation

This application note demonstrates accurate, reproducible quantification of endogenous CRP levels in plasma using a generic kit-based approach.

Application of High- and Low-resolution Mass Spectrometers Combined with Nanoscale LC and Microfluidic LC Platforms for the Quantitative Analysis of Peptide Biomarkers in Human Serum

This application note presents the comprehensive comparison of both tandem quadrupole and time-of-flight mass spectrometers for the quantitative MRM-based analysis of peptide biomarkers.

The microfluidic LC platform combined with the Xevo TQ-S tandem quadrupole mass spectrometer is the ideal LC-MS system for translational research studies where low level quantitation of tryptic peptides is required.

Improvements in Sensitivity for Biotherapeutics Using a Tandem Quadrupole MS

Presented at ASMS 2016, this poster presents data that was generated using a prototype tandem quadrupole MS. Oxytocin, Vancomycin (cyclic peptides), and a large monoclonal antibody were selected as candidate molecules to cover a cross-section of analytically-challenging biotherapeutics.

Microflow LC-MS Application Notebook

This applications notebook contains application notes and white papers featuring ionKey/MS and the iKey Separation device.

See how we are bringing microflow LC-MS to the masses by creating a technology that standardizes and simplifies microflow. These application notes and whitepapers show how to increase sensitivity up to 40X, while reducing sample requirements, and saving up to 60X on solvents - which increases your confidence, saves you time and resources.

Large Molecule Analysis by LC-MS – Survey Infographic

Sponsored by Waters, the Bioanalysis Zone Spotlight has produced an in-depth infographic on large molecule analysis by LC-MS. This infographic displays the results of a community survey, where contributors were asked about their large molecule assays and their opinions on key issues.

Quantification of Intact Protein Biotherapeutics by a Microfluidic LC-MS Platform Poster

This poster aims to evaluate the analytical performance of the ionKey/Xevo G2-XS system for quantification of protein therapeutics at the intact level.

Acquiring and Processing Full Scan or Tof-MRM (Targeted HRMS) Datasets for Quantitative Assays

HRMS data for quantitative applications are becoming more and more widely accepted. UNIFI software enables collection of multiple modes of data and flexible quantitation workflows. UNIFI enables users to develop, process, and analyze HRMS bioanalysis assays with workflows that fit traditional tandem discovery or regulated bioanalytical workflows.

Peptide Separations Application Notebook

This document is a compilation of published LC and LC-MS applications for the analyses of peptides.

Quantitation of Biopharmaceuticals by Mass Spectrometry

The quantitation of proteins in complex biological samples is an important and challenging task. Quantitation of small molecules, while less challenging, also requires some special considerations. This new e-book presents essential information about these analyses, from general precepts through specific considerations when developing an LC-MS-MS method.

Peptide & Protein Bioanalysis Infographic

WHY is it harder to quantify large molecules? WHY are biotherapeutics more important than ever? Check out this infographic which addresses these biologic bioanalysis questions.

Using Microflow LC-MS to Achieve High Sensitivity Analysis of Intact Antibodies

Results from an intact IgG1 mAb mass analysis were used to illustrate sensitivity capabilities that can help biopharm labs extend precious samples.

Fast, Sensitive Quantification of Infliximab from Rat Plasma Using Single, Universal Prep Method

As more drug development focuses on large molecules, traditional “small molecule” scientists find themselves challenged by the complexity and the multitude of potential workflows that exist for protein quantification by LC-MS. This app note describes a simple, standardized approach to protein quantification, where samples are ready for LC-MS analysis in 4–6 hours, and a detection limit of 10 ng/mL for infliximab is achieved.

A Universal, Optimized SPE Protocol for Clean-up of Tryptic Peptides in Protein Bioanalysis

A simple 96-well μElution format enabled clean-up and concentration of digest samples in <20 minutes, without the need for evaporation and reconstitution, preserving low levels of precious tryptic peptides.

Achieving 15-30 Fold Improvement in Sensitivity Utilizing Microfluidics for Peptide Quantification

Critical bioanalytical assays were adapted to integrated microscale LC to reduce sample volumes while increasing sensitivity. Assays for teriparatide, glucagon and human insulin and five analogs were transferred from 2.1 mm analytical scale LC to a 150 µm scale system.

High Resolution Mass Spectrometry Quantitation Explained: Adding Sensitivity and Selectivity Through Instrument Design and Practice

Benchmark studies of the Xevo G2-XS QTof suggests it is >1000 times more sensitive than those introduced just 10 years ago. This jump in improvement can be attributed to both StepWave ion guide technology and a new collision cell design that minimizes background noise, improves ion focusing, and reduces losses in ion transfer.

Quantifying Peptide Biomarkers: Comparing Integrated Microflow LC-MS with Nanoscale LC-MS

Eight LC/MS platforms were examined to identify which combinations of systems delivered the best throughput, sensitivity, linearity, and reproducibility.

Peptide Bioanalysis Using HRMS with Target Enhancement MRM

Increasing demand for protein/peptide bioanalysis by LC-MS techniques has led to a renewed interest in HRMS platforms for use in quantitation because they enable both a larger mass range and additional options for selectivity. The coupling of targeted HRMS with microfluidics offers a powerful, simple workflow for HRMS peptide bioanalysis.

LC-MS Bioanalysis of Proteins via the Surrogate Peptide Approach

Dr. Erin Chambers discusses challenges in workflow options for the bioanalysis of proteins using the surrogate peptide approach; antibodies and the options for signature peptides; the effect of protein- and peptide-level clean up; the quantification of ADCs; and other biologic bioanalysis applications.

An Ultrasensitive Method for the Quantitation of Active and Inactive GLP-1 in Human Plasma

An ultrasensitive, highly-selective immunoaffinity LC-MS/MS (IA LC-MS/MS) assay capable of quantitating endogenous levels of active (7–36 amide) and inactive (9–36 amide) GLP-1 in human plasma was developed. The sensitivity of this assay is better than another LC-MS/MS GLP-1 assay previously reported and many commercially available immunoassays.

Quantification of the Antibody Drug Conjugate (ADC), in Plasma Using a Generic Kit-Based Approach

This application note describes the total mAb quantification of the ADC, ado-trastuzumab emtansine, and the mAb, trastuzumab from rat plasma using the ProteinWorks eXpress Direct Digest Kit and Protocol.

High Throughput Analysis with Microfluidics Under Gradient Conditions

The ionKey/MS System with Xevo G2-XS Qtof HRMS is a high-performance platform that can be used for highly sensitive detection of small molecules, peptides, and proteins. This application shows the ionKey/MS can also be operated under high throughput conditions with a 3-minute full gradient and a 7 μL/min flow rate.

Development of High Sensitivity Intact Monoclonal Antibody (mAb) Analysis Using an Intergrated Microfluidics LC-MS System

LC-MS has emerged as a powerful and robust tool for the characterization of intact monoclonal antibodies (mAb). This approach can also be applied to heterogeneous therapeutic monoclonal antibodies, antibody drug conjugates (ADCs), bispecific mAbs, antibody-antigen complexes and antibody mixtures. The ability to also separate the mAb from impurities, small molecules, dissociated light and heavy chains, allows for cleaner spectra with less ion adducts and the ability to discern glycoform variants.

Reaching New Heights in Targeted Quantitation Using HRMS

Newer technologies are allowing scientists to move beyond traditional small molecules, peptides, and oligonucleotides to proteins, monoclononal antibodies, and ADCs. Three technologies are reviewed: microfluidics, HRMS, and software.