Evaluation of size exclusion chromatography columns packed with sub-3 μm particles for the analysis of biopharmaceutical proteins

Goyon, Alexandre; Beck, Alain; Colas, Olivier; Sandra, Koen; Guillarme, Davy; Fekete, Szabolcs

doi:10.1016/j.chroma.2016.11.056

Cited by 67 publications

(53 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Size-exclusion chromatography (SEC) is a traditional technique widely used for protein characterization and quality control [53]. The main application of SEC is the quantitative determination of protein aggregates.…”

Section: Size-exclusion Chromatographymentioning

confidence: 99%

“…SEC separation should be performed in conditions as inert as possible. Recently, SEC columns packed with sub-3 m particles have been introduced, making possible to perform ultra-high performance size-exclusion (SE-UHPLC or UHP-SEC) separations within 5-10 min analysis time range [53,59,62]. UHP-SEC columns perform two or three times lower plate heights compared to conventional 5-10 m SEC packing.…”

Section: Size-exclusion Chromatographymentioning

confidence: 99%

See 1 more Smart Citation

Current possibilities of liquid chromatography for the characterization of antibody-drug conjugates

Bobály

Fleury-Souverain²,

Beck

et al. 2018

Journal of Pharmaceutical and Biomedical Analysis

Self Cite

View full text Add to dashboard Cite

Antibody Drug Conjugates (ADCs) are innovative biopharmaceuticals gaining increasing attention over the last two decades. The concept of ADCs lead to new therapy approaches in numerous oncological indications as well in infectious diseases. Currently, around 60 CECs are in clinical trials indicating the expanding importance of this class of protein therapeutics. ADCs show unprecedented intrinsic heterogeneity and address new quality attributes which have to be assessed. Liquid chromatography is one of the most frequently used analytical method for the characterization of ADCs. This review summarizes recent results in the chromatographic characterization of ADCs and supposed to provide a general overview on the possibilities and limitations of current approaches for the evaluation of drug load distribution, determination of average drug to antibody ratio (DAR), and for the analysis of process/storage related impurities. Hydrophobic interaction chromatography (HIC), reversed phase liquid chromatography (RPLC), size exclusion chromatography (SEC) and multidimensional separations are discussed focusing on the analysis of marketed ADCs. Fundamentals and aspects of method development are illustrated with applications for each technique. Future perspectives in hydrophilic interaction chromatography (HILIC), HIC, SEC and ion exchange chromatography (IEX) are also discussed.

show abstract

Section: Size-exclusion Chromatographymentioning

confidence: 99%

Section: Size-exclusion Chromatographymentioning

confidence: 99%

Current possibilities of liquid chromatography for the characterization of antibody-drug conjugates

Bobály

Fleury-Souverain²,

Beck

et al. 2018

Journal of Pharmaceutical and Biomedical Analysis

Self Cite

View full text Add to dashboard Cite

show abstract

“…The evaluation of SEC columns packed with sub‐3 and sub‐2 μm particles for the analysis of mAbs and ADCs was described by Fekete et al. showing that an additional gain in time can be achieved without compromising analysis time . The same group also reported the risk of forming on‐column aggregates when applying small‐particle columns under high‐pressure conditions .…”

Section: Native Lc Modesmentioning

confidence: 99%

“…Furthermore, it was demonstrated that a factor of 2 in analysis time could be gained when using 3 μm SEC resins instead of 5 μm particles, and optimizing the column length-to-particlediameter ratio, such that the column efficiency was maintained [31]. The evaluation of SEC columns packed with sub-3 and sub-2 μm particles for the analysis of mAbs and ADCs was described by Fekete et al showing that an additional gain in time can be achieved without compromising analysis time [32,33]. The same group also reported the risk of forming on-column aggregates when applying small-particle columns under high-pressure conditions [34].…”

Section: Aqueous Size-exclusion Chromatography For the Analysis Of Prmentioning

confidence: 99%

Advances in native high‐performance liquid chromatography and intact mass spectrometry for the characterization of biopharmaceutical products

Tassi

Vos

Chatterjee

et al. 2017

J of Separation Science

View full text Add to dashboard Cite

The characterization of biotherapeutics represents a major analytical challenge. This review discusses the current state‐of‐the‐art in analytical technologies to profile biopharma products under native conditions, i.e., the protein three dimensional conformation is maintained during liquid chromatographic analysis. Native liquid‐chromatographic modes that are discussed include aqueous size‐exclusion chromatography, hydrophobic interaction chromatography, and ion‐exchange chromatography. Infusion conditions and the possibilities and limitations to hyphenate native liquid chromatography to mass spectrometry are discussed. Furthermore, the applicability of native liquid‐chromatography methods and intact mass spectrometry analysis for the characterization of monoclonal antibodies and antibody–drug conjugates is discussed.

show abstract

“…Fractions exhibiting antioxidant properties were confirmed by electrophoresis and mass spectrometric analysis to have a molecular mass below 3 kDa. It must be noted that the performance of an SE‐HPLC system in peptide isolation and purification, with concomitant determination of molecular weight, is affected by parameters such as the pore volume, nature of mobile phase, ionic strength, and type of the SEC solid phase matrix (Bouvier & Koza, ; Goyon et al, ). These parameters are essential to minimize unwanted hydrophobic peptide–peptide interactions, and peptide–solid phase interaction; which could potentially alter their molecular weight, yield, and biological properties (Irvine, ).…”

Section: Structure‐informed Separation Processesmentioning

confidence: 99%

Structure-informed separation of bioactive peptides

et al. 2019

View full text Add to dashboard Cite

The application of proteomic and peptidomic technologies for food-derived bioactive peptides is an emerging field in food sciences. These technologies include the use of separation tools coupled to a high-resolution spectrometric and bioinformatic tools for prediction, identification, sequencing, and characterization of peptides. To a large extent, one-dimensional separation technologies have been extensively used as a continuous tool under different optimized conditions for the identification and analysis of food peptides. However, most one-dimensional separation technologies are fraught with significant bottlenecks such as insufficient sensitivity and specificity limits for complex samples. To address this limitation, separation systems based on orthogonal, multidimensional principles, which allow for the coupling of more than one analytical separation tool with different operational principles, provide a higher separation power than one-dimensional separation tools. This review describes the structure-informed separation and purification of protein hydrolyzates to obtain peptides with desirable bioactivities. Practical applicationsApplication of bioactive peptides in the formulation of functional foods, nutraceuticals, and therapeutic agents have increasingly gained scholarly and industrial attention. The bioactive peptides exist originally in protein sources and are only active after hydrolysis of the parent protein. Currently, several tools can be configured in one-dimensional or multidimensional systems for the separation and purification of protein hydrolyzates. The separations are informed by the structural properties such as the molecular weight, charge, hydrophobicity or hydrophilicity, and the solubility of peptides. This review provides a concise discussion on the commonly used analytical tools, their configurations, advantages and challenges in peptide separation. Emphasis is placed on how the structural properties of peptides assist in the separation and purification processes and the concomitant effect of the separation on peptide bioactivity. K E Y W O R D S bioactive peptides, multidimensional separation, peptide purification, separation systems, structure-property relationships [Correction added on 30 January 2019: this article has been added to the issue after an inadvertent omission.]

show abstract

Evaluation of size exclusion chromatography columns packed with sub-3 μm particles for the analysis of biopharmaceutical proteins

Cited by 67 publications

References 26 publications

Current possibilities of liquid chromatography for the characterization of antibody-drug conjugates

Current possibilities of liquid chromatography for the characterization of antibody-drug conjugates

Advances in native high‐performance liquid chromatography and intact mass spectrometry for the characterization of biopharmaceutical products

Structure-informed separation of bioactive peptides

Contact Info

Product

Resources

About