Two-Dimensional Liquid Chromatography (2D-LC): Analysis of Size-Based Heterogeneities in Monoclonal Antibody–Based Biotherapeutic Products

Bhattacharya, Sanghati; Sarin, Deepika

doi:10.56530/lcgc.na.cz9881a2

Cited by 4 publications

(3 citation statements)

References 33 publications

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“…Results show that in either configuration, all protein mixtures were adequately resolved, both the larger (IgM and IgG) and smaller proteins (RNase A, myoglobin, lysozyme, and carbonic anhydrase) (Figure ). This observation is consistent with previous reports that either column arrangement can work effectively . No major peak broadening was observed in each case, suggesting that connecting SEC columns with different pore sizes is an effective way to extend the separation landscape in SEC.…”

Section: Resultssupporting

confidence: 92%

“…Over the past decade, two-dimensional liquid chromatography (2D-LC) has been used to increase peak capacity and expand column selectivity. − This technique has been used to couple orthogonal modes of chromatography, for example, hydrophobic interaction chromatography and reversed-phase liquid chromatography (HIC-RPLC), , ion-exchange chromatography and RPLC (IEC-RPLC), , RPLC and hydrophilic interaction liquid chromatography (RPLC-HILIC) and HIC-SEC, and achiral and chiral separations. ,− 2D-LC has also been used to combine columns operated in the same chromatography mode to increase peak capacity and selectivity, such as IEC-IEC and RPLC-RPLC . More commonly, for biotherapeutic characterization, 2D-LC has been used to couple mass spectrometry (MS)-incompatible LC methods to MS via solvent exchanging between the first and second dimensions .…”

Section: Introductionmentioning

confidence: 99%

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Two-Dimensional SEC-SEC-UV-MALS-dRI Workflow for Streamlined Analysis and Characterization of Biopharmaceuticals

Barrientos,

Singh,

Ukaegbu

et al. 2024

Anal. Chem.

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Section: Resultssupporting

confidence: 92%

Section: Introductionmentioning

confidence: 99%

Two-Dimensional SEC-SEC-UV-MALS-dRI Workflow for Streamlined Analysis and Characterization of Biopharmaceuticals

Barrientos,

Singh,

Ukaegbu

et al. 2024

Anal. Chem.

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“…So far, the combination of SEC and RP for separating complex samples, commonly known as two-dimensional liquid chromatography (2D-LC), has been a trend. By accessing SEC in the first dimension and RP in the second dimension, researchers have successfully analyzed the protein aggregates [58], monoclonal antibodies [59], and pharmaceutical drug oligomers [60]. The separation mechanism of SEC is mainly based on molecular size, while RP is based on hydrophobicity.…”

Section: Matrix Effect Evaluationmentioning

confidence: 99%

An Online-SPE/SEC/LCMS Method for the Detection of N-Nitrosamine Disinfection Byproducts in Wastewater Plant Tailwater

Tang

Chen

et al. 2022

Water

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N-nitrosamines have recently attracted attention as a class of disinfection byproducts and are also a hot spot in environmental studies. Current N-nitrosamine analytical methods typically involve manual solid phase extraction (SPE) of samples followed by quantitative analysis using liquid chromatography-mass spectrometry (LCMS), which is time-consuming and may also fail to eliminate complex matrix effects. Size exclusion chromatography (SEC) is a technique that can separate compounds according to their molecular size. For the first time, this study developed an Online-SPE/SEC/LCMS quantitative analysis method to detect and analyze nine common N-nitrosamine disinfection byproducts in wastewater plant tailwater, including N-dimethylnitrosamine (NDMA) and N-nitrosodiethylamine (NDEA), etc. The samples of 1.0 mL can be directly injected after the simple 0.22 μm membrane filtration. This method reports the combination of SPE, SEC, and RP C18 columns to achieve several functions in a processing time of 20 min, including online enrichment, desalination, and matrix separation for the first time. The method provides good linearity (R2 > 0.999), recoveries ranging from 91.67% to 105.88%, relative standard deviation (RSD) lower than 4.17%, and the limits of detection (LOD) are 0.12–6.60 ng/L. This method alleviates tedious human labor and can effectively overcome the matrix effect (ME < 20%). This method allows for the accurate quantitative analysis of N-nitrosamines with high compatibility in wastewater plant tailwater, rivers, and lakes with a high background matrix. Interested researchers can also use this method as a reference in the online analysis of other specific pollutants after necessary optimization. It can also be utilized for non-targeted screening and targeted analysis of contaminants in water with a wide range of applications, giving valuable information for environmental monitoring.

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Applications of Metabolomics for the Elucidation of Abiotic Stress Tolerance in Plants: A Special Focus on Osmotic Stress and Heavy Metal Toxicity

Mashabela

Masamba

Kappo

2023

Plants

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Plants undergo metabolic perturbations under various abiotic stress conditions; due to their sessile nature, the metabolic network of plants requires continuous reconfigurations in response to environmental stimuli to maintain homeostasis and combat stress. The comprehensive analysis of these metabolic features will thus give an overview of plant metabolic responses and strategies applied to mitigate the deleterious effects of stress conditions at a biochemical level. In recent years, the adoption of metabolomics studies has gained significant attention due to the growing technological advances in analytical biochemistry (plant metabolomics). The complexity of the plant biochemical landscape requires sophisticated, advanced analytical methods. As such, technological advancements in the field of metabolomics have been realized, aided much by the development and refinement of separatory techniques, including liquid and gas chromatography (LC and GC), often hyphenated to state-of-the-art detection instruments such as mass spectrometry (MS) or nuclear resonance magnetic (NMR) spectroscopy. Significant advances and developments in these techniques are briefly highlighted in this review. The enormous progress made thus far also comes with the dawn of the Internet of Things (IoT) and technology housed in machine learning (ML)-based computational tools for data acquisition, mining, and analysis in the 4IR era allowing for broader metabolic coverage and biological interpretation of the cellular status of plants under varying environmental conditions. Thus, scientists can paint a holistic and comprehensive roadmap and predictive models for metabolite-guided crop improvement. The current review outlines the application of metabolomics and related technological advances in elucidating plant responses to abiotic stress, mainly focusing on heavy metal toxicity and subsequent osmotic stress tolerance.

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Two-Dimensional Liquid Chromatography (2D-LC): Analysis of Size-Based Heterogeneities in Monoclonal Antibody–Based Biotherapeutic Products

Cited by 4 publications

References 33 publications

Two-Dimensional SEC-SEC-UV-MALS-dRI Workflow for Streamlined Analysis and Characterization of Biopharmaceuticals

Two-Dimensional SEC-SEC-UV-MALS-dRI Workflow for Streamlined Analysis and Characterization of Biopharmaceuticals

An Online-SPE/SEC/LCMS Method for the Detection of N-Nitrosamine Disinfection Byproducts in Wastewater Plant Tailwater

Applications of Metabolomics for the Elucidation of Abiotic Stress Tolerance in Plants: A Special Focus on Osmotic Stress and Heavy Metal Toxicity

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