The Chromatography Forum

of Delaware Valley

CFDV Symposium 2026

SAVE THE DATE: April 16, 2026

Affiliated Short Course: April 15, 2026

More information on short course available soon.

Student & Young Professional Poster Session: Details available here.

Preliminary Schedule:

Preliminary Speaker List

Frank Zuo

Merck & Co., Inc.

Analytical Challenges in mRNA and Circular RNA Integrity Analysis for Biopharmaceutical Development

The COVID-19 pandemic has highlighted the significance of mRNA technology, which has played a pivotal role in developing vaccines and potentially saving lives. However, this technology faces challenges in manufacturing and analytical control. One crucial challenge is ensuring the integrity of mRNA drug substances and products through thorough analysis for release and stability testing.

There are two widely used analytical techniques for RNA integrity analysis: Capillary Gel Electrophoresis (CGE) and Ion-Pairing Reversed Phase Liquid Chromatography (IP-RPLC). These technologies play a pivotal role in characterizing mRNA materials but are not without limitations. CGE offers the capability to analyze intact mRNA and smaller fragments, yet it falls short when it comes to distinguishing intact species with adduct of mRNA and cationic lipids. On the other hand, IP-RPLC excels in resolving adduct species from intact mRNA, but it grapples with notorious robustness issues, such as repeatability and transferability due to shallow organic gradient. Furthermore, with the advent of circular RNA technology, the challenge extends to separating circular RNA species from their linear counterparts using either of these technologies.

This presentation delves into the analytical development challenges associated with both linear RNA and circular RNA. The methods discussed encompass slab gel electrophoresis, capillary gel electrophoresis, and ion-pairing chromatography, shedding light on the solutions to the analytical challenges of RNA integrity analysis.

Martin Gilar

Waters Corporation

Applications of High-Performance Affinity Chromatography

Advanced therapies increasingly focus on biotherapeutics. The expansion into biopolymer drugs necessitates the development of analytical tools with precise selectivity. Affinity liquid chromatography (aLC) is such technique, useful specific isolation of target analytes from complex media. We describe development of high-performance aLC columns based on direct immobilization (protein A) or biotinylated affinity selectors immobilization on streptavidin sorbent. In the latter case the biotinylated affinity ligands were immobilized and characterized in chromatographic experimental setup. Both proteins and nucleic acids were immobilized on the aLC sorbent and used as affinity selectors.

We describe applications of aLC for selective and sensitive analysis of monoclonal antibodies (mAb), messenger RNA (mRNA) and other nucleic acids. The selective analysis can be performed with exceptional speed and sensitivity using 20 x 2.1 mm column format packed with small nonporous particles. aLC was integrated with size-exclusion chromatography for advanced characterization of unbound and bound aLC fractions.

Jeff Roberts

GSK

SEC Approaches for Novel mAb Biotherapeutics

Size exclusion chromatography (SEC) is the established method for characterizing aggregation and fragmentation of therapeutic monoclonal antibodies (mAbs), but the emergence of engineered constructs—such as bispecifics and antibody–drug conjugates (ADCs)—with increased hydrophobicity has exposed limitations of conventional aqueous SEC conditions. To address this, we developed a rapid, robust SEC method approach for a broad range of mAb formats by incorporating acetonitrile as an organic modifier and using statistical design-of-experiments to define the operating design space. The resulting method delivered improved chromatographic performance for higher hydrophobicity mAbs and constructs without compromising separations of more conventional mAbs; design space and robustness testing across representative molecules established resilient operating ranges for key parameters, and formal validation demonstrated suitability for routine aggregate and fragment analysis across diverse mAb constructs.

Tom Brettell

Professor Emeritus, Cedar Crest College

Method Development in Gas Chromatography

This presentation will outline and discuss the various steps involved in gas chromatography (GC) method development. When developing an effective GC method business decisions such as cost factors and available instrumentation need to be considered. Time also needs to be budgeted for method development. You must first start with a comprehensive evaluation of the sample’s properties and the analytical goals. The method development process includes selecting the ideal chromatographic conditions, instrumentation, and analytical strategies. It requires systematically optimizing parameters such as carrier gas, inlet, column, and detector to improve analytical performance and streamline workflows. Essential steps involve assessing the physicochemical characteristics of the sample, preparing the sample, choosing suitable chromatographic modes and detectors, optimizing the method, and validating its performance. By adhering to this structured process, GC methods can be customized in a timely manner to provide accurate, precise, and dependable results for a wide range of analytical applications.

Mary Ellen McNally

FMC Corporation

Supercritical Fluid Chromatography: Theory and Practical Method Development

Following an overview of fundamental theory and key definitions, this presentation will review the phase diagrams that define the distinctive behavior of supercritical fluids and illustrate why these materials occupy a unique region between liquids and gases. Building on this foundation, the discussion will examine the characteristics of supercritical fluid mobile phases and the physicochemical properties—such as enhanced diffusivity, and reduced viscosity—that can be leveraged to achieve efficient chromatographic separations. Clear distinctions will be drawn between enhanced fluidity chromatography and traditional supercritical fluid chromatography (SFC), emphasizing how each technique utilizes pressure and mobile-phase composition differently. The presentation will also explore how operational parameters including pressure, temperature, density, and modifier concentration independently and collectively influence retention, selectivity, and overall separation performance. By integrating these concepts, attendees will gain a comprehensive understanding of how to optimize SFC methods for a wide range of analytical challenges.

Joe Foley

Drexel University

Capillary Electrophoresis: Theory, Methodology, and Method Development

Following a brief overview of the family of voltage-driven techniques comprised within capillary electrophoresis—zone electrophoresis (CZE), micellar electrokinetic chromatography (MEKC), gel electrophoresis (CGE), isoelectric focusing (CIEF), and isotachophoresis (CITP)—the theory, advantages, and limitations of CZE (a.k.a. "CE") and MEKC are described. The methodology for CE and MEKC is summarized, including sample introduction, analyte focusing (stacking), separation, and detection. A few sets of alternative experimental conditions that address some specific limitations are shared. An approach to method development for CE and MEKC is then presented. Selected applications of small- and large-molecule analyses are described, including the separation of (i) inorganic cations and anions, (ii) enantiomers of amino acids, (iii) charge-variant analysis of antibody-drug conjugates (ADC's), and (iv) a compendium method for the analysis of erythropoietin variants.