Based on an episode of the Concentrating on Chromatography podcast hosted by David Oliva, General Manager of Organomation
Biopharmaceuticals have transformed modern medicine. Monoclonal antibodies, antibody-drug conjugates (ADCs), bispecific antibodies, GLP-1 agonists, gene therapies, and lipid nanoparticle delivery systems are redefining how diseases are treated. But as these therapies become more sophisticated, the analytical challenges surrounding quality assurance and quality control (QA/QC) become dramatically more complex.
On a recent episode of Concentrating on Chromatography, David Oliva spoke with Colette Quinn, Senior Director within the Biologics Business at Waters Corporation, about why biologics QC is fundamentally different from traditional small molecule pharmaceutical analysis and how advanced analytical technologies are reshaping the future of biopharma quality control.
For laboratories performing LC-MS workflows, chromatography-based separations, protein characterization, or sample preparation for biologics analysis, the conversation highlighted many emerging challenges and opportunities directly impacting analytical scientists today.
The Core Difference: Biologics Are Inherently Heterogeneous
One of the most important themes from the discussion was the idea that biologics are not single, static molecular entities. Unlike small molecule drugs, biologics exist as dynamic populations of molecules.
“With small molecules, quality is often about confirming identity and purity against very well-defined specifications,” Quinn explained during the interview. “With biologics, quality is about managing variability.”
This distinction fundamentally changes the role of QA/QC laboratories.
Small molecule pharmaceuticals are relatively straightforward from an analytical perspective. Chemists know the expected molecular structure, molecular weight, stereochemistry, and impurity profile. Traditional chromatographic and spectrometric techniques can often confirm identity with a high degree of certainty.
Biologics are entirely different.
Protein therapeutics can contain:
- Charge variants
- Glycosylation heterogeneity
- Aggregates
- High molecular weight species
- Higher-order structural changes
- Partial unfolding
- Oxidation or deamidation products
Even batches manufactured under highly controlled conditions may display subtle variability. As Quinn noted, “the product is the process” in biologics manufacturing.
This creates enormous analytical demands for QA/QC laboratories.
Why Chromatography Alone Is Sometimes Not Enough
Chromatography remains foundational in biologics QC workflows. Techniques such as:
- Size exclusion chromatography (SEC)
- Ion exchange chromatography (IEX)
- Reverse phase LC
- Affinity chromatography
- Two-dimensional LC (2D-LC)
all play critical roles in monitoring critical quality attributes (CQAs).
According to Quinn, LC addresses “80–90% of the attributes required in biosimilars and innovator therapeutics.”
However, one of the major challenges discussed in the episode is that chromatographic separations can sometimes hide underlying complexity.
The Problem of Co-Eluting Species
A recurring issue in biologics analysis is co-elution.
Analysts may observe:
- Slight shoulders on chromatographic peaks
- Peak broadening
- Increased full width at half maximum (FWHM)
- Subtle retention time shifts
These can indicate heterogeneity within the sample.
But the problem becomes even more complicated when multiple species elute simultaneously.
As Quinn explained, a contaminant with a completely different molecular weight may still elute at the same time as the target protein because of similarities in hydrodynamic radius or higher-order structure.
This creates significant analytical risk.
A chromatogram may appear clean while hidden contaminants or aggregates remain undetected beneath the primary peak.
For QA/QC labs, this can become a major concern because regulatory decisions, release testing, and patient safety may depend on correctly identifying those hidden species.
Orthogonal Analytical Techniques Are Becoming Essential
One of the strongest messages from the interview was the growing importance of orthogonal analytical measurements.
Chromatography alone often cannot provide complete molecular characterization.
This is why techniques such as:
- Multi-angle light scattering (MALS)
- High-resolution mass spectrometry (HRMS)
- Dynamic light scattering (DLS)
- Differential scanning calorimetry (DSC)
- Capillary electrophoresis (CE)
are increasingly being integrated into QC environments.
Why MALS Matters
Quinn described several real-world examples where light scattering techniques revealed contaminants that traditional SEC methods missed.
In one case, a partially unfolded contaminant co-eluted with the target molecule on an SEC column. The contaminant had a completely different molecular weight but appeared chromatographically identical.
Light scattering measurements ultimately identified the problem.
This allowed the laboratory to:
- Detect the contaminating species
- Adjust purification strategies
- Improve product quality
These examples demonstrate why relying exclusively on retention time assumptions can be dangerous in biologics analysis.
Understanding High Molecular Weight Species
Another particularly challenging area involves high molecular weight species (HMWS).
Many biologics contain:
- Dimers
- Oligomers
- Large aggregates
Differentiating between these structures is critically important because aggregates can impact:
- Efficacy
- Pharmacokinetics
- Immunogenicity
- Stability
Quinn emphasized that QC laboratories increasingly need more than simple percentage quantification.
They must determine:
- What the species actually are
- Whether they are reversible oligomers or irreversible aggregates
- How they evolve over time
This is especially relevant for high-profile biologics such as GLP-1 agonists and antibody-drug conjugates.
The Rise of 2D-LC in Biologics Analysis
One especially interesting topic from the podcast was the growing use of two-dimensional liquid chromatography (2D-LC).
As biologics become more structurally complex, traditional one-dimensional separations are often insufficient.
This is particularly true for:
- Bispecific antibodies
- Multispecific antibodies
- Antibody-drug conjugates (ADCs)
These molecules frequently contain both hydrophobic and hydrophilic regions, making them difficult to fully characterize using conventional methods.
2D-LC enables analysts to combine multiple separation modes into a single workflow.
For example:
- Affinity chromatography in the first dimension
- SEC or reverse phase LC in the second dimension
This dramatically improves separation power and characterization depth.
As biologics continue evolving, multidimensional separations are likely to become increasingly important in both development and QC laboratories.
Automation and the “Lights Out Lab”
Beyond instrumentation, the discussion also explored automation and digital transformation within QC environments.
Automation in QA/QC has historically focused on:
- Sample handling
- Pipetting
- System suitability testing
- Automated reporting
But Quinn believes software integration and data analysis remain underutilized.
Modern QC labs generate enormous datasets across multiple analytical platforms:
- LC systems
- Mass spectrometers
- Light scattering instruments
- Environmental monitoring systems
- Particle analysis tools
Yet these datasets often remain siloed.
The future likely involves centralized data ecosystems capable of integrating:
- Instrument outputs
- Environmental variables
- Historical trends
- Predictive analytics
- Machine learning models
This broader connectivity may ultimately enable:
- Real-time release testing
- Improved trending analysis
- Reduced false investigations
- Better process understanding
Quinn also discussed the concept of the “lights out lab,” where automated workflows and intelligent systems reduce human intervention while increasing consistency and reproducibility.
Why Method Transfer to QC Is So Difficult
Another key challenge discussed in the episode is method transfer.
Methods developed in R&D environments often do not translate easily into QC labs.
Development methods may prioritize:
- Deep characterization
- Flexibility
- Exploratory analysis
QC methods require:
- Robustness
- Simplicity
- Reproducibility
- Regulatory compliance
According to Quinn, transferring methods into QC can become a six-month process involving:
- Validation
- Platform compatibility testing
- Software integration
- Qualification procedures
This is why “quality by design” (QbD) and method lifecycle management are increasingly important.
Analytical methods must be developed from the beginning with QC implementation in mind.
Data Integrity Is Becoming Central to QC
One of Quinn’s strongest points was that data integrity remains the number one priority in QC environments.
Modern biologics QA/QC is no longer simply about running assays.
It requires:
- Understanding variability
- Interpreting complex datasets
- Recognizing hidden risks
- Making defensible release decisions
As regulatory expectations evolve, software systems capable of:
- Automated comparisons
- Audit trails
- Trending analysis
- Cross-platform integration
will become increasingly valuable.
This trend also aligns with recent regulatory interest in advanced analytics and alternative testing approaches.
In 2025 and 2026, the FDA has signaled growing interest in reducing reliance on animal testing while expanding the use of advanced analytical characterization and predictive modeling in pharmaceutical development.
What This Means for Analytical Laboratories
For analytical laboratories supporting LC-MS, protein characterization, and biologics workflows, the implications are significant.
As biologics become more sophisticated:
- Orthogonal measurements will become increasingly necessary
- Data interpretation will matter as much as instrumentation
- Advanced characterization techniques will move downstream into QC
- Software and workflow integration will become critical
- Multidimensional chromatography will continue expanding
Importantly, these trends also increase the importance of high-quality sample preparation workflows.
Techniques such as nitrogen blowdown evaporation remain essential for:
- Protein and peptide sample concentration
- Extraction cleanup
- LC-MS preparation
- Trace impurity analysis
As QC workflows become more analytically demanding, consistency during sample preparation becomes even more important for reproducibility and regulatory confidence.
The Future of Biologics QC
Perhaps the most important takeaway from the interview was that biologics QA/QC is evolving from a reactive compliance function into a proactive scientific discipline.
QC laboratories are no longer simply verifying specifications after manufacturing.
They are increasingly:
- Monitoring product trends
- Detecting emerging risks
- Understanding molecular behavior
- Enabling process optimization
- Supporting patient safety through advanced analytics
And as biologics continue increasing in complexity, analytical science will remain at the center of that transformation.