Comprehensive solutions for characterization, confirmation, and monitoring of your vaccine

Protein-based vaccines are composed of purified or recombinant proteinaceous antigens from a pathogen. These include virus-like particle (VLP) vaccines that are proteins self-assembling into a particle that resembles a virus. Subunit vaccines typically need adjuvants to boost immune response (see Vaccine Formulation and Process Residuals for adjuvant-specific analytics). Recombinant proteins are grown in cells in bioreactors, purified, and then formulated. For vaccines to be acceptable to regulatory agencies, in-depth analytical characterization must be performed and purity/impurity criteria set (CQAs) to ensure product safety and efficacy.

Key applications in vaccine chararcterization

  • Higher Order Structural Analysis
  • Intact Protein Analysis Workflows
  • Protein Aggregate Analysis
  • Robust Charge Variant Profiling
  • Complete Solutions for Glycan Profiling
  • Intact Glycoform Analysis
  • Glycopeptide Workflow
  • Peptide Mapping
  • Residual Host Cell Protein Analysis (HCPs)
  • Peptide Mapping for QC/Lot release
  • Release Glycan Monitoring

Characterize the full diversity of your vaccine biomolecules

Get high resolution, high efficiency separation of your vaccine associated biological samples. Our unique column chemistries for biological samples have been specifically designed for the separation of proteins, peptides, monoclonal antibodies, carbohydrates, nucleic acids, and more.

Chromatography Consumables Resources and Information
Select the Correct Chemistry for Your Analysis

Accelerate your vaccine development with confidence

Thermo Scientific Vanquish HPLC and UHPLC systems are the most advanced liquid chromatography instruments available. The Vanquish LC platform improves performance, repeatability, and dependability with no trade-offs in quality or ease-of-use. Vanquish LC systems provide the versatility to solve the toughest vaccine analytics challenges.

Ensuring the safety of your final vaccine products

A highly flexible Thermo Scientific Dionex ion chromatography (IC) system provides customers with the freedom to develop, explore, and run different methods simultaneously. Utilizing unique IC technologies with unparalleled robustness and capabilities, one can accurately identify and monitor formulation and process residual components and ensure appropriate adjuvant concentrations, with ease.

Universal detection and quantification for non-chromophore containing adjuvants and formulations

Utilize the increased separation speed and resolution of the Thermo Scientific Vanquish UHPLC system with charged aerosol detection (CAD). CAD can detect and quantify virtually any substance with enhanced linear dynamic range and accurate, consistent response. Measure low-level impurities and active pharmaceutical ingredients (APIs) simultaneously, as well as other compounds, regardless of chemical structure using CAD.

Comprehensive vaccine characterization

Thermo Scientific Orbitrap Exploris mass spectrometry system is quantitative high-resolution, accurate-mass (HRAM) liquid chromatography mass spectrometry (LC-MS) with record-setting performance for your next-generation of vaccine products. Extraordinary power is now available in an expanded portfolio to meet your exact analytical requirements, deliver comprehensive characterization of your most advanced vaccine drug products, and accelerate their development.

Track everything, simplify compliance, and boost productivity

Achieve productivity and compliance with one software platform. As an integral part of your laboratory's IT structure, the latest Thermo Scientific Chromeleon 7.3 Chromatography Data System (CDS) is vital for compliant, efficient, and reliable handling of today’s chromatography and mass spectrometry data. It ensures data quality and manages all the analytical processes from instrument control, to raw data storage and processing, to generating the final results.

Characterize the full diversity of your vaccine biomolecules

Get high resolution, high efficiency separation of your vaccine associated biological samples. Our unique column chemistries for biological samples have been specifically designed for the separation of proteins, peptides, monoclonal antibodies, carbohydrates, nucleic acids, and more.

Chromatography Consumables Resources and Information
Select the Correct Chemistry for Your Analysis

Accelerate your vaccine development with confidence

Thermo Scientific Vanquish HPLC and UHPLC systems are the most advanced liquid chromatography instruments available. The Vanquish LC platform improves performance, repeatability, and dependability with no trade-offs in quality or ease-of-use. Vanquish LC systems provide the versatility to solve the toughest vaccine analytics challenges.

Ensuring the safety of your final vaccine products

A highly flexible Thermo Scientific Dionex ion chromatography (IC) system provides customers with the freedom to develop, explore, and run different methods simultaneously. Utilizing unique IC technologies with unparalleled robustness and capabilities, one can accurately identify and monitor formulation and process residual components and ensure appropriate adjuvant concentrations, with ease.

Universal detection and quantification for non-chromophore containing adjuvants and formulations

Utilize the increased separation speed and resolution of the Thermo Scientific Vanquish UHPLC system with charged aerosol detection (CAD). CAD can detect and quantify virtually any substance with enhanced linear dynamic range and accurate, consistent response. Measure low-level impurities and active pharmaceutical ingredients (APIs) simultaneously, as well as other compounds, regardless of chemical structure using CAD.

Comprehensive vaccine characterization

Thermo Scientific Orbitrap Exploris mass spectrometry system is quantitative high-resolution, accurate-mass (HRAM) liquid chromatography mass spectrometry (LC-MS) with record-setting performance for your next-generation of vaccine products. Extraordinary power is now available in an expanded portfolio to meet your exact analytical requirements, deliver comprehensive characterization of your most advanced vaccine drug products, and accelerate their development.

Track everything, simplify compliance, and boost productivity

Achieve productivity and compliance with one software platform. As an integral part of your laboratory's IT structure, the latest Thermo Scientific Chromeleon 7.3 Chromatography Data System (CDS) is vital for compliant, efficient, and reliable handling of today’s chromatography and mass spectrometry data. It ensures data quality and manages all the analytical processes from instrument control, to raw data storage and processing, to generating the final results.

Select the application below to view full product characterization approaches for protein-based vaccine analysis:

Hydrogen Deuterium Exchange (HDX)

Orbitrap-based HDX MS using the Thermo Scientific Orbitrap Eclipse Tribrid MS offers a robust method for the analysis of protein conformation, conformation dynamics, and protein-protein interactions. Full structural characterization is critical in protein-based vaccine development where the subtle but critical local conformational changes can impact safety and efficacy.

Hydrogen Deuterium Exchange (HDX)

Intact mass analysis

Among the analytical tools used for the characterization of proteins-based vaccines, high-resolution accurate-mass (HRAM) mass spectrometry has become increasingly important for quantification of the various glycoforms and protein variant analysis. New methods coupling UHPLC with Orbitrap-based mass spectrometers give rapid insight into the proteoform profile for direct comparison of protein based vaccine products during discovery, development, and QC.

Intact Mass Analysis

Aggregation of proteinaceous antigens can result in an incorrect vaccine dosage and undesired immune response. Consequently, monitoring protein aggregates is important for safety and quality assurance.

Protein Aggregate Analysis

Size exclusion chromatography (SEC) is the most commonly used method for protein aggregate analysis. In SEC, sample molecules are passed through a column containing porous polymer or silica beads to facilitate separation of species based on their size.

Software designed for routine GMP protein aggregation analysis

Perform your protein aggregation assays in an enterprise environment—from method creation to final reporting. Streamline your laboratory workflow using Chromeleon 7.2 Chromatography Data System (CDS) software. This software delivers superior instrument control, automation, and data processing for compliant GMP biopharmaceutical manufacturing and QA/QC environments.

The presence of the charged state can significantly impact the structure and stability of your protein-based vaccine. It is therefore necessary to understand the charge profile of the protein vaccine and accurately characterize any microheterogeneity.

Robust Charge Variant Profiling

Our next generation cation exchange column extends column lifetime and method robustness - the Thermo Scientific MAbPac SCX-10 LC Column is for high-resolution, high efficiency analysis of therapeutic proteins and associated variants. The unique nonporous pellicular resin provides the resolving power for therapeutic protein variants. A hydrophilic layer surrounds polymeric beads and eliminates hydrophobic interactions with the resin, resulting in very efficient peaks. A proprietary grafted cation-exchange surface provides pH selectivity control for high-resolution separations.

Run your routine HPLC, UHPLC, or LC-MS protein charge variant analyses in an enterprise environment—from method creation to final reporting—using Chromatography data solution for GMP testing of protein charge variants.

Biotherapeutic glycoproteins with complex glycosylation patterns have the potential to easily fall out of specification with changes in biomanufacturing processes. To meet regulatory demands manufacturers must carefully characterize glycosylation of proteins and its relation to the clinical activity. The complete analysis of a glycoprotein provides information on the primary structure of the oligosaccharides as well as their variation at individual glycosylation sites.

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Answer the question – how many glycoforms are in my product?

Perform rapid analysis of protein glycosylation at the intact level. Determine intact glycoform composition quickly and simply to make process-level decisions on the fly. Combine rapid separations with accurate glycoprotein mass spectrometry (MS) for confidence in process-related glycosylation differences.

Intact Glycoform Analysis

Answer the question - where do the glycans reside in my product?

Analogous to standard peptide mapping analysis workflows, glycopeptide analysis can yield glycosylation site information with advanced LC-MS/MS fragmentation techniques such as electron transfer dissociation (ETD) and higher energy C-trap dissociation (HCD) using Thermo Scientific Orbitrap-based mass spectrometry. Learn more: Fragmentation versatility for complete glycopeptide characterization

Glycopeptide Workflow

Determine the exact location of glycosylation with advanced fragmentation techniques on the Thermo Scientific Orbitrap Eclipse Tribrid mass spectrometer, which enables multiple fragmentation techniques (CID, HCD, ETD) combined with ionization spectra (such as EThcD) and product dependent acquisition (such as HCDpdETD), to generate a pair of HCD and ETD spectra. At any stage of MSn, fragment ion detection in either the ion trap or Orbitrap mass analyser offers a new level of versatility and performance for the most challenging applications, such as the identification and location of glycopeptides.

From protein-based vaccine discovery and development, through the clinical testing phase and on to bioproduction and QA/QC, peptide mapping remains a key analytical methodology.

Peptide Mapping

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Application information: Protein Digestion for Mass Spectrometry

Product information: Orbitrap Exploris 480 Mass Spectrometer

HCPs are low-level, process-related protein impurities in protein-based vaccines products derived from the host organism during manufacturing. During expression of a recombinant proteinaceous antigen, host cell systems can express many endogenous proteins. Purification of the antigen from these HCP contaminants can be challenging, with low-level contamination remaining after purification. Host cell proteins (HCPs) can affect efficacy and cause unwanted immunogenicity. Detection and quantification of residual HCPs as potential process-related impurities is critical for biopharmaceutical companies in accordance with regulatory agency guidelines.

Residual Host Cell Protein Analysis (HCPs)
 Orbitrap Exploris 480 Mass SpectrometerOrbitrap Exploris 240 Mass Spectrometer
 Orbitrap Exploris 480 Mass SpectrometerOrbitrap Exploris 240 Mass Spectrometer
AdvantagesExtraordinary simplified within a compact footprint where next level exploration delivers quantifiable accuracy and the selectivity required to comprehensively identify and quantify host cell protein contaminants.Realize complete BioPharma characterization and identify, quantify and confirm more host cell proteins rapidly and with confidence.
Resolving power480,000 at m/z 195Up to 240,000 at m/z 200
Scan speedUp to 40Hz22Hz
Mass range40 to 6,000 m/z (up to 8,000 m/z with the BioPharma option)4 to 8,000 m/z
Dynamic range>5000:1>5000:1
Mass accuracyInternal <1 ppm RMS; External: <3 ppm RMSInternal <1 ppm RMS; External: <3 ppm RMS

Peptide mapping methods are often developed and evaluated with combined UV and MS detection, to simplify the transfer to routine environments where UV detection is used alone. In high-throughput workflows, peptide mapping experiments are performed for protein identity confirmation, PTM characterization, and stability studies.

Monitoring and quality control (QC) methods: Peptide Mapping for QC/Lot release

Glycans are often derivatized prior to analysis, but it is also possible and convenient particularly for routine environments to analyze both N-glycans and O-Glycans in a label-free form, typically using ion chromatography (IC) or liquid chromatography with charged aerosol detection.

Release glycan monitoring

IC technique is sensitive and selective and resolves glycans according to charge, size, composition, isomers and linkage.

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Thermo Scientific Vanquish Flex UHPLC

Product information: Compliant chromatography software essential for all the monitoring and quality control methods mentioned above

Product information

Sign up to attend workshops of interest

  • BioPharma Finder software for characterization of protein and oligonucleotide biotherapeutics
  • BioPharma Finder software for in-depth protein characterization, intact mass analysis, peptide mapping, and new host cell protein workflow
  • BioPharma Finder software for in-depth oligonucleotide characterization, intact mass analysis, oligonucleotide analysis and mRNA mapping
  • GlycanExplorer software for high throughput biotherapeutic glycan characterization
  • Proteome Discoverer software for increasing proteoform level insight with quantitative top-down proteomics

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