Bio-conjugates

Bio-conjugates

Comprehensively characterize the complex, heterogeneous structures of bio-conjugates (including antibody drug conjugates) with Waters fit-for-purpose sample preparation, columns, UPLC, QTof MS, DLS, MALS, and informatics solutions. Waters is committed to supporting the development of safe and effective therapies for cancers and other diseases by helping you generate accurate ADC profiles to characterize and monitor critical quality attributes.

Comprehensively characterize the complex, heterogeneous structures of bio-conjugates (including antibody drug conjugates) with Waters fit-for-purpose sample preparation, columns, UPLC, QTof MS, DLS, MALS, and informatics solutions. Waters is committed to supporting the development of safe and effective therapies for cancers and other diseases by helping you generate accurate ADC profiles to characterize and monitor critical quality attributes.

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

ADCs are a class of biopharmaceutical drugs that deliver cancer-killing agents directly to cancer cells – a far more targeted approach than conventional chemotherapy. Optimizing ADCs requires analytical technologies to comprehensively characterize the molecule, including its biochemical attributes like drug-to-antibody ratio (DAR) and sites of conjugation.

As biopharmaceutical organizations continue to explore the use of ADCs as a class of cancer therapies, they find significant challenges with analyzing their complex, heterogenous structures. However, a well-characterized ADC is critical to advance from discovery through clinical development and to commercialization.

Successfully address your characterization challenges with exceptional insight into your molecules from Waters fit-for-purpose ultra-performance liquid chromatography (UPLC), high resolution mass spectrometry (MS), column chemistries, and informatics solutions. Trust Waters to help simplify complex molecule analysis and accelerate your ADC characterization and monitoring of critical quality attributes.


Applications

ultraDAWN™ real-time MALS directly monitors product attributes post-purification, enhancing yield and flexibility. It connects downstream purification and enrichment methods, such as IEX, FPLC, or TFF, to measure molecular weight, concentration, size, and impurities\impurities in real-time, streamlining processing and maximizing yield.

ultraDAWN™ real-time MALS directly monitors product attributes post-purification, enhancing yield and flexibility. It connects downstream purification and enrichment methods, such as IEX, FPLC, or TFF, to measure molecular weight, concentration, size, and impurities\impurities in real-time, streamlining processing and maximizing yield.


Application Notes

Application Notes

The efficacy of bioconjugates like ADCs greatly depends on the binding affinity and stability of the monoclonal antibody (mAb) portion of the molecule. Light scattering techniques and calorimetry can narrow down antibody candidates, optimized for binding kinetics well as stability.

The efficacy of bioconjugates like ADCs greatly depends on the binding affinity and stability of the monoclonal antibody (mAb) portion of the molecule. Light scattering techniques and calorimetry can narrow down antibody candidates, optimized for binding kinetics well as stability.


Application Notes

Application Notes

Whether coupled to mass spectrometry, MALS, or as a standalone measurement, separating structural features of ADCs is critical to our enhanced understanding during product development and release. Waters chemistry solutions continue to provide innovative approaches to offer ultimate chromatographic performance and robustness.

Whether coupled to mass spectrometry, MALS, or as a standalone measurement, separating structural features of ADCs is critical to our enhanced understanding during product development and release. Waters chemistry solutions continue to provide innovative approaches to offer ultimate chromatographic performance and robustness.


Application Notes

Application Notes

Mass spectrometry and light scattering techniques provide insight into multiple quality attributes of ADCs from early to late stages in the development process. These attributes include determining optimal conjugated payload ratio (for example, quantifying Drug to Antibody Ratio), sulfide-bond mapping, characterizing post-translational modifications, and identifying conjugate positional isomers. Waters has developed workflows and applications to support these critical analyses.

Mass spectrometry and light scattering techniques provide insight into multiple quality attributes of ADCs from early to late stages in the development process. These attributes include determining optimal conjugated payload ratio (for example, quantifying Drug to Antibody Ratio), sulfide-bond mapping, characterizing post-translational modifications, and identifying conjugate positional isomers. Waters has developed workflows and applications to support these critical analyses.


Application Notes

Application Notes

Conjugated protein therapeutics often face stability and aggregation challenges due chemical modifications and incorporation of hydrophobic drugs. High-throughput DLS enables rapid screening and stability studies, while SEC-MALS offers detailed characterization in formulation development.

Conjugated protein therapeutics often face stability and aggregation challenges due chemical modifications and incorporation of hydrophobic drugs. High-throughput DLS enables rapid screening and stability studies, while SEC-MALS offers detailed characterization in formulation development.


Application Notes

Application Notes


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

To quickly move an ADC-based therapeutic out of development and into the clinic, Igenica Biotherapeutics wanted industry-standard instrumentation to analyze the complexity and variation within the ADC. This case study describes how Waters solutions, including Waters Biopharmaceutical Platform Solution with UNIFI, provided a superior window of insight into the molecules being developed by Igenica.

To quickly move an ADC-based therapeutic out of development and into the clinic, Igenica Biotherapeutics wanted industry-standard instrumentation to analyze the complexity and variation within the ADC. This case study describes how Waters solutions, including Waters Biopharmaceutical Platform Solution with UNIFI, provided a superior window of insight into the molecules being developed by Igenica.


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.

Designed specifically for biopharmaceutical routine monitoring

Designed specifically for biopharmaceutical routine monitoring

Obtain greater simplicity, accuracy, and reproducibility for attribute monitoring 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

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, without time-consuming tasks like 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 antibody drug conjugate (ADC) separation challenges with ACQUITY UPLC Systems with 2D LC Technology, a family of systems and columns that offer the lowest dispersion for maximum peak capacity.

  • 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 biopharma LC-MS applications, intact mass, peptide mapping, peptide MAM, and released N-glycan analysis.

  • 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

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.
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).
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



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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