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Mass Spectrometry: Complex Analysis01:21

Mass Spectrometry: Complex Analysis

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Mass spectrometry is an important technique for the identification of pure compounds. However, it has some limitations for the analysis of complex mixtures, often due to excessive fragmentation making the spectrum too complicated to decipher. Mass spectrometry can be combined with suitable separation methods in sequence, forming hyphenated methods, which are useful in the analysis of complex mixtures.
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The mass analyzer is a crucial component of the mass spectrometer. In the ionization chamber, the vaporized sample is bombarded with a high-energy electron beam to generate a radical cation and further fragment into neutral molecules, radicals, and cations. A series of negatively charged accelerator plates accelerate the cations into the mass analyzer. The mass analyzer separates ions according to their mass-to-charge (m/z) ratios and then directs them to the detector. The common types of mass...
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Mass spectrometry is an analytical technique used to determine the molecular mass and molecular formula of a compound. The basic principle of mass spectrometry is to generate ions from the analyte molecule and measure these ion abundances against their molecular mass.  One common type of ionization, known as electrospray ionization or EI, bombards the analyte molecules in the gas phase with high-energy electron beams. The electron beams displace an electron from the molecule and leave...
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This lesson details the instrumentation of a mass spectrometer—a physical instrument to perform mass spectrometry on analyte molecules and record the characteristic mass spectra. This is achieved via three chief functions:
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Multi attribute method implementation using a High Resolution Mass Spectrometry platform: From sample preparation to

Sofia B Carvalho1,2, Ricardo A Gomes1,2, Anja Pfenninger3

  • 1iBET, Instituto de Biologia Experimental e Tecnologica, Oeiras, Portugal.

Plos One
|January 27, 2022
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Summary
This summary is machine-generated.

The multi-attribute method (MAM) using high-resolution mass spectrometry offers efficient quality control for biopharmaceuticals like monoclonal antibodies (mAbs). This method streamlines analysis of critical quality attributes, reducing time and costs.

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Area of Science:

  • Biopharmaceutical quality control
  • Analytical chemistry
  • Mass spectrometry

Background:

  • Biopharmaceutical quality control is increasingly complex due to evolving regulatory demands and intricate product structures.
  • Traditional methods for assessing product quality attributes (PQAs) are often time-consuming and costly.
  • Mass Spectrometry (MS)-based methods, particularly the multi-attribute method (MAM), are emerging as powerful tools for comprehensive biopharmaceutical characterization.

Purpose of the Study:

  • To describe the implementation process of a Multi-Attribute Method (MAM) using a QTOF high-resolution mass spectrometry platform.
  • To demonstrate the capability of MAM for simultaneous monitoring of various Product Quality Attributes (PQAs) in monoclonal antibodies (mAbs).
  • To discuss troubleshooting for sample preparation and LC-MS parameters for high-throughput batch analysis.

Main Methods:

  • Implementation of a MAM workflow on a QTOF high-resolution mass spectrometry platform.
  • Utilized NIST mAb reference material and an in-process mAb sample for method validation.
  • Employed high mass accuracy/high-resolution MS data for direct and simultaneous monitoring of PQAs.

Main Results:

  • Successfully monitored critical PQAs including glycosylation profiles, methionine oxidation, tryptophan dioxidation, asparagine deamidation, N-terminal pyro-glutamic acid formation, and glycation.
  • The MAM workflow proved effective for comprehensive characterization of a downstream processing polishing platform.
  • Demonstrated the utility of MAM in a comparability study following technology transfer between laboratories.

Conclusions:

  • The described MAM implementation provides a robust and efficient workflow for biopharmaceutical quality control.
  • MAM offers a streamlined approach, replacing multiple orthogonal methods and reducing analysis time and costs.
  • This high-resolution MS-based MAM serves as a valuable reference analytical tool for biopharmaceutical characterization and comparability studies.