Bioconjugate Development

Bioconjugate Development

Comprehensively characterize complex, heterogeneous bio‑conjugated molecules with Waters fit‑for‑purpose sample preparation, columns, UPLC, MS, DLS, MALS, and informatics solutions. Our platform supports bioconjugate development from discovery to commercialization—helping you monitor critical quality attributes (CQAs), including conjugation ratio and distribution, with speed and confidence.

Comprehensively characterize complex, heterogeneous bio‑conjugated molecules with Waters fit‑for‑purpose sample preparation, columns, UPLC, MS, DLS, MALS, and informatics solutions. Our platform supports bioconjugate development from discovery to commercialization—helping you monitor critical quality attributes (CQAs), including conjugation ratio and distribution, with speed and confidence.

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

Overview

Bioconjugate development without specialized expertise

Bioconjugates are increasingly central to next-generation therapeutics, enabling the precise delivery of potent payloads through targeted molecular design. Optimizing complex, heterogeneous molecules requires robust analytical technologies to fully characterize critical attributes such as conjugation sites, payload distribution, and overall molecular heterogeneity.

As bioconjugated molecules advance from discovery to commercialization, their structural diversity and complexity present significant analytical challenges. Comprehensive characterization is essential to ensure safety, efficacy, and consistent product quality.

Waters provides fit-for-purpose solutions to deliver deep insight into bioconjugate structure and quality. Simplify your complex molecule analysis and accelerate the characterization and monitoring of critical quality attributes, regardless of your bioconjugate structure.


Applications

Mass spectrometry and light scattering deliver multi‑attribute insight across early and late stages of bioconjugate development. Determine optimal conjugated payload ratio (quantify DAR/conjugation ratio), perform disulfide‑bond mapping, assess post‑translational modifications, and identify positional isomers. Waters provides proven workflows for high‑confidence ADC characterization and broader bioconjugate analytics.

Mass spectrometry and light scattering deliver multi‑attribute insight across early and late stages of bioconjugate development. Determine optimal conjugated payload ratio (quantify DAR/conjugation ratio), perform disulfide‑bond mapping, assess post‑translational modifications, and identify positional isomers. Waters provides proven workflows for high‑confidence ADC characterization and broader bioconjugate analytics.


Application Notes

Application Notes
Acquire, analyze and manage mass spectrometry information in the clinical laboratory with MassLynx IVD Mass Spectrometry Software, and increase the speed at which you can convert your sample data into valuable knowledge.

Directly monitor product attributes post‑purification to enhance yield and flexibility. ultraDAWN™ real‑time MALS/DLS connects to downstream workflows (IEX, FPLC, TFF) to measure molecular weight, concentration, size, and impurities in real time—streamlining processing and maximizing yield across bioconjugate development and scale‑up.

Directly monitor product attributes post‑purification to enhance yield and flexibility. ultraDAWN™ real‑time MALS/DLS connects to downstream workflows (IEX, FPLC, TFF) to measure molecular weight, concentration, size, and impurities in real time—streamlining processing and maximizing yield across bioconjugate development and scale‑up.


Application Notes

Application Notes
Scientist holding a test tube next to a bioreactor

Bioconjugate performance hinges on mAb binding strength and stability. High‑throughput light scattering and calorimetry enable rapid assessment of aggregation, solution stability, binding kinetics, and thermal robustness—helping prioritize candidates with durable target engagement for bioconjugate development.

Bioconjugate performance hinges on mAb binding strength and stability. High‑throughput light scattering and calorimetry enable rapid assessment of aggregation, solution stability, binding kinetics, and thermal robustness—helping prioritize candidates with durable target engagement for bioconjugate development.


Application Notes

Application Notes
Pipette depositing samples in a well panel

Whether hyphenated to mass spectrometry, coupled to MALS, or with optical detection, chromatographically resolving structural features of conjugated proteins is critical to development and release. Waters LC and chemistry solutions deliver innovative approaches for ultimate performance and robustness—supporting ADC characterization and broader bioconjugate development.

Whether hyphenated to mass spectrometry, coupled to MALS, or with optical detection, chromatographically resolving structural features of conjugated proteins is critical to development and release. Waters LC and chemistry solutions deliver innovative approaches for ultimate performance and robustness—supporting ADC characterization and broader bioconjugate development.


Application Notes

Application Notes
Empower Method Validation Manager (MVM) provides chromatography labs with a powerful workflow-based solution for automated chromatographic method validation, from protocol planning through final reporting.

Conjugated protein therapeutics face stability and aggregation risks due to chemical modifications and hydrophobic payloads, with aggregates spanning nanometers to microns. High‑throughput DLS enables rapid screening, SEC‑MALS provides detailed size and conjugation analysis, and advanced backgrounded membrane imaging (BMI) differentiates intrinsic and extrinsic sub‑visible particulates—delivering end‑to‑end characterization to de‑risk formulation and process development in bioconjugate development programs.

Conjugated protein therapeutics face stability and aggregation risks due to chemical modifications and hydrophobic payloads, with aggregates spanning nanometers to microns. High‑throughput DLS enables rapid screening, SEC‑MALS provides detailed size and conjugation analysis, and advanced backgrounded membrane imaging (BMI) differentiates intrinsic and extrinsic sub‑visible particulates—delivering end‑to‑end characterization to de‑risk formulation and process development in bioconjugate development programs.


Application Notes

Application Notes
Scientific researcher in a lab looking at vial


Case Study: How Waters is Helping to Support the Development of Novel Therapies

To rapidly advance an ADC therapeutic into the clinic, Igenica Biotherapeutics required industry‑standard instrumentation to analyze ADC complexity and variability. This case study shows how Waters solutions delivered a superior window of insight—accelerating decisions in ADC characterization and bioconjugate development.

To rapidly advance an ADC therapeutic into the clinic, Igenica Biotherapeutics required industry‑standard instrumentation to analyze ADC complexity and variability. This case study shows how Waters solutions delivered a superior window of insight—accelerating decisions in ADC characterization and bioconjugate development.


Closeup of vaccination vial and syringe

Solutions


With Waters systems, characterize complex bio-conjugates in less time and with greater confidence.

With Waters systems, characterize complex bio-conjugates in less time and with greater confidence.

Purposeful design for bio-conjugate characterization and monitoring

Purposeful design for bio-conjugate characterization and monitoring

Obtain greater accuracy, reproducibility and product insight from attribute monitoring, conjugation ratio, and drug distribution analysis of bio-conjugates with the BioAccord LC-MS System.

  • Real-Time Process Monitoring
  • Candidate Screening
  • Chromatography
  • Identity and Post-Translational Modification Analysis
  • Protein-Conjugate Ratio
  • Aggregation and Stability

Complete coverage for confident characterization

Complete coverage for confident characterization

Maximize sample information for your analytes from detailed characterization to accurate quantitation with the Xevo G3 QTof, which enables complete confidence in your bio-conjugates analytical data.

  • Real-Time Process Monitoring
  • Candidate Screening
  • Chromatography
  • Identity and Post-Translational Modification Analysis
  • Protein-Conjugate Ratio
  • Aggregation and Stability

Delivering performance and speed without compromise

Delivering performance and speed without compromise

Multi-reflecting time-of-flight technology delivers mass resolution consistent over a broad m/z range independent of scan speed, even at the fastest acquisition rates. Unlock sub-ppm mass accuracy for confident identification and greater productivity in your lab

  • Identity and Post-Translational Modification Analysis
  • Aggregation and Stability
  • Candidate Screening

The universal platform for liquid chromatography

The universal platform for liquid chromatography

Eliminate unpredictability of analyte losses due to metal interactions with MaxPeak Premier Solutions, a portfolio of LC, LC-MS, and column chemistries designed to get maximum limits of detection for challenging compounds, while minimizing time-consuming system passivation.

  • Real-Time Process Monitoring
  • Candidate Screening
  • Chromatography
  • Identity and Post-Translational Modification Analysis
  • Protein-Conjugate Ratio
  • Aggregation and Stability

The lowest dispersion system for maximum peak capacity

The lowest dispersion system for maximum peak capacity

Solve your complex UPLC/UHPLC bioconjugate separation challenges with ACQUITY UPLC Systems with 2D LC Technology, a family of systems optimized for powerful separation of complex molecules

  • Real-Time Process Monitoring
  • Candidate Screening
  • Chromatography
  • Identity and Post-Translational Modification Analysis
  • Protein-Conjugate Ratio
  • Aggregation and Stability

Uniquely versatile detector for every developmental stage

Uniquely versatile detector for every developmental stage

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

  • Candidate Screening
  • Chromatography
  • Identity and Post-Translational Modification Analysis
  • Protein-Conjugate Ratio
  • Aggregation and Stability

Measure size and zeta potential in native buffer

Measure size and zeta potential in native buffer

The DynaPro™ ZetaStar™ instrument combines DLS, SLS, and ELS in one platform. Designed for biopharma in mind, it can measure zeta-potential in formulation buffer without dilution.

  • Aggregation and Stability
  • Chromatography

High-throughput size and stability screening

High-throughput size and stability screening

Measure size, polydispersity, and Tagg data for hundreds of samples with as little as 4µL per well. The DynaPro™ DLS Plate Reader uses standard-well plates; integrate with any liquid handler and further boost productivity.

  • Candidate Screening
  • Aggregation and Stability

Increase yield and monitor product quality with confidence

Increase yield and monitor product quality with confidence

ultraDAWN™ directly monitors titer, size, and impurities in real-time. It connects downstream purification and enrichment methods, such as IEX, FPLC, or TFF.

  • Real-Time Process Monitoring

Waters software streamlines data acquisition, processing, and reporting ADC analysis results.

Waters software streamlines data acquisition, processing, and reporting ADC analysis results.

Accelerate your biopharmaceutical analysis

Accelerate your biopharmaceutical analysis

Accelerate the journey from sample analysis to decision making using waters_connect for biopharmaceuticals, with tailored workflows designed for bioconjugate analysis with integrated, automated calculation of conjugation ratio and distribution

  • Real-Time Process Monitoring
  • Candidate Screening
  • Chromatography
  • Identity and Post-Translational Modification Analysis
  • Protein-Conjugate Ratio
  • Aggregation and Stability

As biopharmaceutical organizations explore the use of bio-conjugates and ADCs as a class of cancer therapies, significant challenges are found in analyzing their complex, heterogeneous structures. Waters provides columns and consumables for these unique analytical challenges by facilitating assessment of many important Critical Quality Attributes (CQAs).

As biopharmaceutical organizations explore the use of bio-conjugates and ADCs as a class of cancer therapies, significant challenges are found in analyzing their complex, heterogeneous structures. Waters provides columns and consumables for these unique analytical challenges by facilitating assessment of many important Critical Quality Attributes (CQAs).

Hydrophobic Interaction Chromatography (HIC) for DAR determination

Hydrophobic Interaction Chromatography (HIC) for DAR determination

Achieve high-quality LC-based analyses and determine DAR of cysteine-conjugated and highly hydrophobic ADCs using Waters Protein-Pak Hi Res Hydrophobic Interaction Chromatography (HIC) HPLC Columns for fast, efficient separations.

  • Real-Time Process Monitoring
  • Candidate Screening
  • Chromatography
  • Identity and Post-Translational Modification Analysis
  • Protein-Conjugate Ratio
  • Aggregation and Stability

Reversed-phase columns for DAR determination & drug distribution

Reversed-phase columns for DAR determination & drug distribution

Reversed-Phase Columns for bio-conjugates and Antibody Drug Conjugates with MaxPeak HPS such as the ACQUITY and XBridge Premier Protein BEH C4 Columns are uniquely designed to minimize secondary interactions between ADCs and column hardware and suitable for the analysis of lysine-conjugated ADCs. Unlike cysteine-conjugated ADCs, the intra-chain disulfide bonds that maintain linkages between the heavy and light chains of the mAb for the lysine-conjugated ADCs are intact. Therefore, reversed-phase chromatography is suitable for the analysis of lysine-conjugated ADCs when linker chemistry is not labile at acidic pH. In addition, Waters offers Reversed-Phase Columns with solid-core particle technology, such as BioResolve RP mAb Polyphenyl for intact and subunit analysis to determine the localization of linked drugs at the domain level.

  • Candidate Screening
  • Chromatography
  • Identity and Post-Translational Modification Analysis
  • Protein-Conjugate Ratio
  • Aggregation and Stability

Achieve consistent aggregation accuracy with SEC Columns

Achieve consistent aggregation accuracy with SEC Columns

Eliminate the need to create multiple SEC eluent mobile phases with different proteins  by using a platform method with phosphate buffered saline (i.e., PBS) for consistently accurate aggregate, monomer, and fragmented data with Waters SEC Columns. In addition, for bio-conjugates which can be hydrophobic creating unwanted secondary interactions, there is no need to add organic solvent with MaxPeak Premier Protein SEC Columns and it’s PEO bonded BEH particles.

  • Real-Time Process Monitoring
  • Candidate Screening
  • Chromatography
  • Identity and Post-Translational Modification Analysis
  • Protein-Conjugate Ratio
  • Aggregation and Stability

Characterize and monitor charge variants with cation exchange chromatography

Characterize and monitor charge variants with cation exchange chromatography

Accurately measure charge variant profiles of cysteine-linked and lysine-linked ADCs with the BioResolve SCX mAb Column and BioResolve CX pH Buffer Concentrates for salt or pH gradients with minimal secondary interactions in optical detection. For structural insights with mass spectrometry in ion exchange, our MS-compatible buffers, IonHance CX-MS pH Buffer Concentrates, make online IEX-MS a reality with high-purity, volatile buffers suitable for mass spectrometry.

  • Real-Time Process Monitoring
  • Candidate Screening
  • Chromatography
  • Identity and Post-Translational Modification Analysis
  • Protein-Conjugate Ratio
  • Aggregation and Stability

Measure oligonucleotide conjugation with Anion Exchange Columns

Measure oligonucleotide conjugation with Anion Exchange Columns

Antibody oligonucleotide conjugates (AOCs) are commonly used in protein diagnostics and are a promising therapeutic for targeted cell therapy. The net negative charge of an AOC expands with each additional conjugation site, resulting in increased retention on an anion exchange resin. The GenPak FAX Column contains a weak anion exchange sorbent that exhibits controlled retentivity and a unique selectivity for protein and oligonucleotide samples. Recent application work demonstrates that AEX can be coupled with MALS to generate absolute molecular weight information.

  • Real-Time Process Monitoring
  • Candidate Screening
  • Chromatography
  • Identity and Post-Translational Modification Analysis
  • Protein-Conjugate Ratio
  • Aggregation and Stability

Precisely locate glycans within a protein structure with glycoprotein columns

Precisely locate glycans within a protein structure with glycoprotein columns

Hydrophilic Interaction Chromatography (HILIC) has been widely used to separate small polar compounds, yet its application to large biomolecules, other than released glycans, has been surprisingly limited. With Waters widepore Glycoprotein Columns, it is now possible to use HILIC to glean previously unattainable information from intact proteins (with or without glycosylation), protein fragments, and complex, released glycans.

  • Real-Time Process Monitoring
  • Candidate Screening
  • Chromatography
  • Identity and Post-Translational Modification Analysis
  • Protein-Conjugate Ratio
  • Aggregation and Stability

Rapid, automatable, and robust protein digestion workflow

Rapid, automatable, and robust protein digestion workflow

Waters PeptideWorks Tryptic Protein Digestion Kits for manual or automated workflows uniquely provide your lab with high efficiency, reproducible peptide maps in under 2.5 hours—4x faster than average home-brew methods. Achieve 30-minute digestions with a 1:5 enzyme:peptide ratio, 78% reduction in missed cleavages, and a 98% reduction in contaminated autolysis peaks. PeptideWorks is ideal for drug conjugation site determination, protein structural characterization, protein identification, and protein modification monitoring including post-translation modifications.

  • Real-Time Process Monitoring
  • Candidate Screening
  • Chromatography
  • Identity and Post-Translational Modification Analysis
  • Protein-Conjugate Ratio
  • Aggregation and Stability

Glycan analysis just got easier and more sensitive

Glycan analysis just got easier and more sensitive

Re-imagine a streamlined, fast, and highly sensitive way to profile N-glycans on ADCs with the GlycoWorks Kit. Achieve unprecedented fluorescent and mass spectrometric performance for released n-glycan analysis while maximizing productivity with a solution that is automatable on Waters Andrew+ Pipetting Robot. With just 3 easy steps in as little as 30 minutes, the GlycoWorks RapiFluor-MS N-Glycan Kit reduces complicated, time-consuming sample preparation.

  • Real-Time Process Monitoring
  • Candidate Screening
  • Chromatography
  • Identity and Post-Translational Modification Analysis
  • Protein-Conjugate Ratio
  • Aggregation and Stability

Take the complexity out of protein quantification

Take the complexity out of protein quantification

Simplify and accelerate protein digestion with ProteinWorks eXpress Digest Kits. Pre-measured, lot-traceable reagents and standardized protocols for accurate, precise, and sensitive LC-MS quantification of proteins, including ADCs, in plasma and serum. Achieve unmatched performance and flexibility for reproducible protein digestion with kits optimized for various workflows, including protein bioanalysis/DMPK.

  • Real-Time Process Monitoring
  • Candidate Screening
  • Chromatography
  • Identity and Post-Translational Modification Analysis
  • Protein-Conjugate Ratio
  • Aggregation and Stability

Sensitive and robust antibody titer measurements earlier in cell harvest

Sensitive and robust antibody titer measurements earlier in cell harvest

Accelerate antibody titer measurements with Waters BioResolve Protein A Affinity Columns featuring MaxPeak Premier Technology. These high-performance columns help you achieve up to 7x greater sensitivity, enabling faster decision-making and better process control. These columns also unlock advanced analytical workflows, including simplified Direct Connect 2D ProA‑SEC and ProA‑MS methods. Streamline your processes and gain unprecedented, in‑depth insights into your antibodies.

  • Real-Time Process Monitoring
  • Candidate Screening

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

Streamline sample preparation for quantitative LC-MS analyses with verified workflows that help minimize variability, improve traceability, and simplify method transfer.

  • Real-Time Process Monitoring
  • Candidate Screening
  • Chromatography
  • Identity and Post-Translational Modification Analysis
  • Protein-Conjugate Ratio
  • Aggregation and Stability

Your success is just a click away

Your success is just a click away

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

  • Real-Time Process Monitoring
  • Candidate Screening
  • Chromatography
  • Identity and Post-Translational Modification Analysis
  • Protein-Conjugate Ratio
  • Aggregation and Stability

Make science more accessible with Waters Capital

Make science more accessible with Waters Capital

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.

  • Real-Time Process Monitoring
  • Candidate Screening
  • Chromatography
  • Identity and Post-Translational Modification Analysis
  • Protein-Conjugate Ratio
  • Aggregation and Stability

The data speaks for itself

The data speaks for itself
Cysteine conjugated ADC analysis using HIC. Drug distribution was determined for three different cysteine-conjugated ADC samples with increasing drug load

Cysteine conjugated ADC analysis using HIC. Drug distribution was determined for three different cysteine-conjugated ADC samples with increasing drug load.

Gaussian-fitted DSC ADC results collected on the BioAccord System for native (top) and a high DAR mAb (bottom)

Gaussian-fitted DSC ADC results collected on the BioAccord System for native (top) and a high DAR mAb (bottom).

Fast SEC analysis of trastuzumab emtansine (A) and bevacizumab (B) as performed on an XBridge Premier Protein SEC 250 Å 2.5 µm 4.6 x 150 mm column. Mobile phase: 1x PBS, pH 7.4, flow rate F = 0.7 mL/min, temperature T = 25 °C, injected volume 1 μL, detection: 280 nm (UV)

Fast SEC analysis of trastuzumab emtansine (A) and bevacizumab (B) as performed on an XBridge Premier Protein SEC 250 Å 2.5 µm 4.6 x 150 mm column. Mobile phase: 1x PBS, pH 7.4, flow rate F = 0.7 mL/min, temperature T = 25 °C, injected volume 1 μL, detection: 280 nm (UV).

Peptide Mapping (UV chromatograms) comparison of a conjugated and unconjugated Trastuzumab

Peptide Mapping (UV chromatograms) comparison of a conjugated and unconjugated Trastuzumab
Top: Trastuzumab biosimilar tryptic digestion.
Bottom: Trastuzumab emtansine (Kadcyla), antibody drug conjugate tryptic digestion.
Both tryptic digestions utilized RapiZyme Trypsin and PeptideWorks Tryptic Protein Digestion Kit with 100 µL injection (~ 10 µg) on XSelect Premier Peptide CSH C18, 2.5 µm, 2.1 x 150 mm column. Additional peaks observed in Kadcyla chromatogram (bottom) compared to unconjugated trastuzumab (top).


Webinars and Resources



  • eBook

Characterizing Antibody Conjugates Guidebook

Characterizing Antibody Conjugates Guidebook

Related

Rely on Waters as your single source solution for antibody drug conjugate (ADC) chromatography columns, designed to provide an accurate drug-to-antibody ratio for discovery, development, and manufacturing applications.

Learn more about Bio-conjugates Solutions.

Learn more about Bio-conjugates Solutions.

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