Biosimilar Analysis

Biosimilar Analysis

Perform rapid and robust biosimilar analysis of therapeutic candidates with Waters’ advanced analytical technologies. Waters columns, consumables, instrumentation, and informatics enable comprehensive biosimilar characterization and help establish the comparability of a therapeutic drug candidate to its reference product with confidence.

Perform rapid and robust biosimilar analysis of therapeutic candidates with Waters’ advanced analytical technologies. Waters columns, consumables, instrumentation, and informatics enable comprehensive biosimilar characterization and help establish the comparability of a therapeutic drug candidate to its reference product with confidence.
Ensure viral vector quality with Waters LC and MS solutions for characterization and routine analysis.
Ensure viral vector quality with Waters LC and MS solutions for characterization and routine analysis.

Overview

Faster, clearer protein therapeutic answers

Global regulators increasingly expect detailed analytical and biological evidence to demonstrate comparability between biosimilars and reference products. This requires developers to thoroughly assess multiple protein batches, emphasizing precise biosimilar protein characterization and deeper characterization of biotherapeutic proteins.

Waters’ integrated solutions provide a comprehensive platform for streamlined, reliable analysis of biosimilar and reference product attributes, including:

  • Primary structure (sequence)
  • Higher‑order structure, including aggregation
  • Post‑translational modifications
  • Intentional chemical modifications such as PEGylation

Establish biosimilar comparability with confidence—partner with Waters for safer, more effective medicines.

Regulatory Insight: FDA Draft Guidance (2025)

Recent FDA draft guidance enables developers to bypass comparative clinical efficacy studies if advanced analytical data provides sufficient evidence of biosimilarity—potentially eliminating one of the costliest development steps and accelerating timelines dramatically.

This shifts an even greater responsibility onto biologics and biosimilars protein analysis and robust analytical testing.

Read the blog

Applications

Owing to the inherent complexity of biotherapeutics, developers must demonstrate comprehensive molecular characterization, robust and well‑controlled manufacturing processes, and ongoing monitoring of product and critical quality attributes. Within biosimilar development, intact mass and subunit analyses are core analytical tools, used to confirm molecular mass, detect sequence errors and post‑translational modifications, and identify major molecular variants that warrant further investigation.

Owing to the inherent complexity of biotherapeutics, developers must demonstrate comprehensive molecular characterization, robust and well‑controlled manufacturing processes, and ongoing monitoring of product and critical quality attributes. Within biosimilar development, intact mass and subunit analyses are core analytical tools, used to confirm molecular mass, detect sequence errors and post‑translational modifications, and identify major molecular variants that warrant further investigation.

Application Notes

Comprehensive tools for intact protein analysis

Peptide mapping is essential for establishing similarity through high sequence coverage and the detection of clinically relevant modifications at the peptide level. Non‑reducing digests further support characterization of biosimilars by enabling disulfide linkage confirmation and equivalence verification to innovator molecules.

Waters solutions deliver high‑precision data to support faster decision‑making across development and manufacturing.

Peptide mapping is essential for establishing similarity through high sequence coverage and the detection of clinically relevant modifications at the peptide level. Non‑reducing digests further support characterization of biosimilars by enabling disulfide linkage confirmation and equivalence verification to innovator molecules.

Waters solutions deliver high‑precision data to support faster decision‑making across development and manufacturing.

Application Notes

Workflows to streamline comparability studies at the peptide level

Protein higher‑order structure is critical in determining functional equivalence.

Comparing structural profiles to the reference product reveals potential functional differences or confirms acceptable similarity.

Waters’ structural biology solutions integrate precise chromatography, robust sample management, and advanced mass spectrometry for efficient acquisition and interpretation of complex HOS datasets.

HDX‑MS and CIU enable detailed assessment of conformational dynamics and stability—key elements in advanced biosimilar characterization workflows.

Protein higher‑order structure is critical in determining functional equivalence.

Comparing structural profiles to the reference product reveals potential functional differences or confirms acceptable similarity.

Waters’ structural biology solutions integrate precise chromatography, robust sample management, and advanced mass spectrometry for efficient acquisition and interpretation of complex HOS datasets.

HDX‑MS and CIU enable detailed assessment of conformational dynamics and stability—key elements in advanced biosimilar characterization workflows.

Application Notes

Demonstrate comparability of protein higher order structure with HDX-MS

Glycosylation affects efficacy, safety, and immunogenicity.

Understanding glycan patterns is essential for demonstrating equivalent quality, consistency, and manufacturing robustness. Waters offers leading solutions for analysis of glycopeptides and released N‑glycans, providing deep insight into major and minor glycoforms to support comparability and process control.

Glycosylation affects efficacy, safety, and immunogenicity.

Understanding glycan patterns is essential for demonstrating equivalent quality, consistency, and manufacturing robustness. Waters offers leading solutions for analysis of glycopeptides and released N‑glycans, providing deep insight into major and minor glycoforms to support comparability and process control.

Application Notes

Develop robust and comparable processes by monitoring profiles of glycopeptides and released N-glycans

Multi‑Attribute Biosimilarity Assessment (MALS)
Absolute molecular weight determination and high‑resolution aggregation and fragmentation profiling are critical for establishing biosimilarity. The DAWN detector enables confident structural comparability across development, supporting sensitive detection of clinically relevant differences without reliance on reference standards.

Comprehensive Particle Characterization for Biosimilars
Robust visible and subvisible particle characterization supports biosimilar comparability by enabling early risk identification, consistent control strategies, and alignment with regulatory expectations. High‑throughput particle data across development and QC from Aura Particle Analyzers, helps demonstrate equivalence in product quality throughout the biosimilar lifecycle.

Multi‑Attribute Biosimilarity Assessment (MALS)
Absolute molecular weight determination and high‑resolution aggregation and fragmentation profiling are critical for establishing biosimilarity. The DAWN detector enables confident structural comparability across development, supporting sensitive detection of clinically relevant differences without reliance on reference standards.

Comprehensive Particle Characterization for Biosimilars
Robust visible and subvisible particle characterization supports biosimilar comparability by enabling early risk identification, consistent control strategies, and alignment with regulatory expectations. High‑throughput particle data across development and QC from Aura Particle Analyzers, helps demonstrate equivalence in product quality throughout the biosimilar lifecycle.

Application Notes

Syringe and vaccine

Webinar: Deploying Waters Solutions to Enhance Biosimilar Development

Hear how Waters solutions streamline mAb subunit LC‑MS workflows for multiple attribute monitoring (MAM) applied to biosimilar candidates throughout development. Presenter: Similis Bio.

Hear how Waters solutions streamline mAb subunit LC‑MS workflows for multiple attribute monitoring (MAM) applied to biosimilar candidates throughout development. Presenter: Similis Bio.
View Webinar
Preparative separations are purposeful and practical, meant for isolating targets from mixtures.

Solutions

With Waters systems, characterize complex biosimilars in less time and with greater confidence.

With Waters systems, characterize complex biosimilars in less time and with greater confidence.

Efficient product attribute monitoring of biosimilars

Efficient product attribute monitoring of biosimilars
BioAccord LC-MS System

Enable biosimilar comparability analyses in every lab with the Waters BioAccord LC-MS System, and waters_connect software for compliance-ready data analysis and reporting.

  • Intact Mass and Subunit Analysis
  • Peptide Mapping and MAM
  • Glycosylation Analysis
  • Aggregation and Fragment Impurity Analysis

Complete high-resolution capability for confident characterization and method transfer

Complete high-resolution capability for confident characterization and method transfer
Xevo G3 QTof

Accelerate biosimilar characterization and monitoring with the Xevo G3 QTof high-performance HRMS benchtop system, delivering accurate qualitative and quantitative results.

  • Intact Mass and Subunit Analysis
  • Peptide Mapping and MAM
  • Glycosylation Analysis
  • Aggregation and Fragment Impurity Analysis
  • Higher Order Structure (HOS): HDX MS & CIU

Delivering cutting-edge analytical performance

Delivering cutting-edge analytical performance
SYNAPT XS High Resolution Mass Spectrometer

Obtain ultimate flexibility with the SYNAPT XS mass spectrometer that offers greater freedom of analytical choice to support scientific creativity and technical success for analysis of biosimilars.

  • Intact Mass and Subunit Analysis
  • Peptide Mapping and MAM
  • Glycosylation Analysis
  • Aggregation and Fragment Impurity Analysis
  • Higher Order Structure (HOS): HDX MS & CIU

Ultimate power, ultimate performance. Redefine the boundaries of science

Ultimate power, ultimate performance. Redefine the boundaries of science
SELECT SERIES Cyclic IMS

Combine novel cyclic ion mobility separation with state-of-the-art high-performance, time-of-flight mass spectrometry with the SELECT SERIES Cyclic IMS, enabling researchers to reliably identify and monitor biosimilar attributes

  • Intact Mass and Subunit Analysis
  • Peptide Mapping and MAM
  • Glycosylation Analysis
  • Aggregation and Fragment Impurity Analysis
  • Higher Order Structure (HOS): HDX MS & CIU

Uniquely versatile detector for every developmental stage

Uniquely versatile detector for every developmental stage
DAWN

The DAWNTM MALS detector assists at every development stage. Couple with EclipseTM to measure binding affinity during discovery. Place downstream of any LC and perform advance analysis of biosimilar protein-conjugate ratio, post-translational modifications, and aggregation.

  • Intact Mass and Subunit Analysis
  • Peptide Mapping and MAM
  • Glycosylation Analysis

Compliance-ready, networkable applications-based software to streamline LC-MS data acquisition, processing, and reporting.

Compliance-ready, networkable applications-based software to streamline LC-MS data acquisition, processing, and reporting.

Biosimilar analysis workflows

Biosimilar analysis workflows
waters_connect for biopharmaceuticals

Streamline biosimilar analysis workflows and unify LC‑MS systems across organizations with this compliant‑ready, networked informatics platform.

  • Intact Mass and Subunit Analysis
  • Peptide Mapping and MAM
  • Glycosylation Analysis
  • Aggregation and Fragment Impurity Analysis

Achieve high efficiency separations for  the analysis and purification of biosimilars including size-exclusion chromatography for aggregation, reversed-phase for intact, subunit, peptide mapping, ion exchange for charge variants, and hydrophilic chromatography for glycoprotein analysis.

Achieve high efficiency separations for  the analysis and purification of biosimilars including size-exclusion chromatography for aggregation, reversed-phase for intact, subunit, peptide mapping, ion exchange for charge variants, and hydrophilic chromatography for glycoprotein analysis.

Innovative solutions for biosimilar characterizations for charge variant and intact and subunit analyses

Innovative solutions for biosimilar characterizations for charge variant and intact and subunit analyses
BioResolve Columns and Consumables

BioResolve RP mAb Polyphenyl  Columns provide improved peak capacity and minimal carry-over for intact and subunit analysis of proteins and mAbs specifically QC-tested with a NIST-based, mAb subunit standard. BioResolve SCX mAb Columns and Buffers reliably separate charge variants for a wide range of mAbs using optical or LC-MS compatible mobile phases.

  • Intact Mass and Subunit Analysis
  • Peptide Mapping and MAM
  • Glycosylation Analysis

More confident size variant separations for biosimilars

More confident size variant separations for biosimilars
ACQUITY and XBridge Premier Protein SEC 250 Å Columns

Minimize secondary ionic or hydrophobic interactions in size-based separations with ACQUITY and XBridge Premier Protein SEC 250 Å Columns with MaxPeak High Performance Surface (HPS) Technology for reliable protein aggregate, monomer, and fragment analyses using a “platform” type method with 30% increase in resolution.

  • Intact Mass and Subunit Analysis
  • Peptide Mapping and MAM
  • Glycosylation Analysis

Maximize sensitivity while minimizing metal adsorption of acidic deamidated peptides

Maximize sensitivity while minimizing metal adsorption of acidic deamidated peptides
MaxPeak Premier Peptide Columns

Gain enhanced recovery and sensitivity for acidic peptides with Waters ACQUITY, XSelect, and XBridge Premier Peptide Columns that are QC batch-tested with Waters Cytochrome c Digest Standard for increased column-to-column performance consistency.

  • Intact Mass and Subunit Analysis
  • Peptide Mapping and MAM
  • Glycosylation Analysis

Achieve quick, clean and complete protein digestion

Achieve quick, clean and complete protein digestion
RapiZyme Trypsin, Mass Spectrometry (MS) Grade

Achieve quick, clean, and complete mAb and biosimilar digestions in just 30 minutes with RapiZyme Trypsin, Mass Spectrometry (MS) Grade, a highly activie autolysis-resistant enzyme that reduces the likelihood of artificial modifications occurring during sample preparation prior to LC-MS analysis.

  • Peptide Mapping and MAM
  • Protein Digestion
  • Peptide Mapping
  • Tryptic Digestion
  • Glycosylation Analysis

Streamline peptide mapping sample preparation

Streamline peptide mapping sample preparation
PeptideWorks Tryptic Protein Digestion Kits

Achieve high efficiency, reproducible peptide maps in under 2.5 hours with PeptideWorks Tryptic Protein Digestion Kits for manual or automated workflows that are 4x faster than average home-brew methods and provide a 78% reduction in missed cleavages.

  • Peptide Mapping and MAM
  • Protein Digestion
  • Peptide Mapping
  • Tryptic Digestion
  • Glycosylation Analysis

Gain control over your glycan separations

Gain control over your glycan separations
MaxPeak Premier Glycan Columns

Obtain fast, high resolution glycan identification for your biosimilars with Waters complete line of glycan columns to analyze glycoproteins, glycopeptides and released N-glycans with exceptional reproducibility.

  • Intact Mass and Subunit Analysis
  • Peptide Mapping and MAM
  • Glycosylation Analysis

Rapid, sensitive, and easy detection of N-glycans

Rapid, sensitive, and easy detection of N-glycans
GlycoWorks RapiFluor-MS Released N-Glycan Kits

Achieve unprecedented fluorescent and mass spectrometric performance for glycan detection with GlycoWorks N-glycan Kits that improve the throughput of N-glycan sample preparation so you don’t have to compromise between speed and sensitivity.

  • Intact Mass and Subunit Analysis
  • Peptide Mapping and MAM
  • Glycosylation Analysis

Faster, more efficient online protein digestion in HDX-MS workflows

Faster, more efficient online protein digestion in HDX-MS workflows
Enzymate BEH Pepsin Column

Fast, efficient, and reproducible online pepsin digestion is critical in experiments that require more peptide coverage. With the Waters Enzymate BEH Pepsin Column, you can easily incorporate online protein digestion into existing HDX- MS workflows. The Enzymate BEH Pepsin Column efficiently digests intact proteins into peptides in an online HDX system, typically in about 30 seconds.

  • Peptide Mapping and MAM
  • Glycosylation Analysis

Achieve precise titer measurements for your advanced antibody modalities with innovative affinity particles.

Achieve precise titer measurements for your advanced antibody modalities with innovative affinity particles.
BioResolve™ Protein A Affinity Columns

Accelerate antibody titer measurements with BioResolve Protein A Affinity Columns featuring MaxPeak Premier Technology. Achieve up to 7× greater sensitivity, faster decision‑making, and improved process control.

These columns also enable advanced workflows, including Direct Connect 2D ProA‑SEC and ProA‑MS methods.

  • Peptide Mapping and MAM
  • Glycosylation Analysis

Gain efficiency and walk away time with verified lab automation solutions.

Gain efficiency and walk away time with verified lab automation solutions.

Enhance sample prep efficiency and throughput

Enhance sample prep efficiency and throughput
Laboratory Automation and Equipment

Streamline sample preparation for LC-MS analyses with Waters Laboratory Automation and Equipment that deliver verified workflows to help minimize variability, improve traceability, and simplify method transfer.

  • Peptide Mapping and MAM
  • Efficacy and Immunogenicity
  • Glycosylation Analysis

Achieve testing efficiencies and accurate results using streamlined Waters services and support.

Achieve testing efficiencies and accurate results using streamlined Waters services and support.

Your success is just a click away

Your success is just a click away
Waters Global Services

Optimize your laboratory's productivity and success with Waters Global Services to maintain peak system performance, minimize downtime, address application challenges, and support stringent compliance requirements.

  • Peptide Mapping and MAM
  • Intact Mass and Subunit Analysis
  • Glycosylation Analysis

Make science more accessible with Waters Capital

Make science more accessible with Waters Capital
Payment Solutions

Maximize resources and minimize risk with payment options from Waters Capital, including upgrading aging equipment, getting customized support, and bundling services into one monthly payment.

  • Peptide Mapping and MAM
  • Intact Mass and Subunit Analysis
  • Glycosylation Analysis

The data speaks for itself

The data speaks for itself
Peptide MAM: Mirror plots showing the comparison of Inflectra to Remicade for both (left) unstressed samples and (right) samples subjected to two weeks of elevated temperature stress.
Peptide MAM: Mirror plots showing the comparison of Inflectra to Remicade for both (left) unstressed samples and (right) samples subjected to two weeks of elevated temperature stress.
Peptide MAM: Selected product quality attributes for Remicade (infliximab) and three biosimilars subjected to thermal stress, T0, T1 (1 week), and T2 (2 weeks). Injections 1–3=Remicade (infliximab), 4–6=Inflectra (infliximab), 7–9=Avsola (infliximab), and 10–12=Renflexis (infliximab).
Peptide MAM: Selected product quality attributes for Remicade (infliximab) and three biosimilars subjected to thermal stress, T0, T1 (1 week), and T2 (2 weeks). Injections 1–3=Remicade (infliximab), 4–6=Inflectra (infliximab), 7–9=Avsola (infliximab), and 10–12=Renflexis (infliximab).
Mirror plots comparing an innovator infliximab sample (Remicade) to two biosimilar samples, (left) Inflectra and (right) Renflexis. The difference in peak intensity is shown as an overlay on each plot, and the Venn diagrams indicate the number of identified glycoforms unique to each sample and those present in both samples
Mirror plots comparing an innovator infliximab sample (Remicade) to two biosimilar samples, (left) Inflectra and (right) Renflexis. The difference in peak intensity is shown as an overlay on each plot, and the Venn diagrams indicate the number of identified glycoforms unique to each sample and those present in both samples.
Overlaid TIC chromatograms of an infliximab originator (Remicade) and a biosimilar (Renflexis) released N-glycan analysis. Some of the glycans differing between the two samples, either in presence or relative abundance, are indicated in the chromatogram with their structures
Overlaid TIC chromatograms of an infliximab originator (Remicade) and a biosimilar (Renflexis) released N-glycan analysis. Some of the glycans differing between the two samples, either in presence or relative abundance, are indicated in the chromatogram with their structures.
Stacked plots of SEC separations on selected columns for biosimilar mAb sample using 1X Dulbecco’s phosphate buffer saline as a mobile phase are shown.
Stacked plots of SEC separations on selected columns for biosimilar mAb sample using 1X Dulbecco’s phosphate buffer saline as a mobile phase are shown.

Webinars and Resources

  • Blog

How Analytical Technologies Support the Development of Biosimilar Drugs

How Analytical Technologies Support the Development of Biosimilar Drugs
Download
  • Brochure

BioAccord LC-MS System with ACQUITY Premier

BioAccord LC-MS System with ACQUITY Premier
Download
  • Infographic

Protein and Peptide Attribute Monitoring Workflows

Protein and Peptide Attribute Monitoring Workflows
Download
  • On Demand Webinar

Access expert webinars on biosimilar development and analytical chromatography. 

Access expert webinars on biosimilar development and analytical chromatography. 
Download

Related

Intact Protein Analysis
Protein molecular weight must be quickly confirmed, along with glycoform heterogeneity determination. Waters technologies deliver quick, accurate protein molecular weights for confirmation and determination of intact proteins.

Peptide Mapping
Achieve unmatched resolution of complicated protein digests, exceptional glycopeptide resolution, and shorter analysis times with UPLC separations and optimally selective and resolving peptide separation chemistries.

Glycan Solutions
Improve the quality of your glycan structure analysis results with Waters complete and easy-to-use workflows for analyzing glycans and glycoproteins.

Hydrogen Deuterium Exchange - Mass Spectrometry
Answers for all of your structural questions for biotherapeutics, complexes, and protein folding/stabillity studies. Waters offers unrivaled instrumentation, chemistries, informatics, and training programs.

Learn more about Biosimilars Solutions.

Learn more about Biosimilars Solutions.
Ensure viral vector quality with Waters LC and MS solutions for characterization and routine analysis.
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