In-capillary (electrophoretic) digestion-reduction-separation: A smart tool for middle-up analysis of mAb.

Comprehensive characterization of physicochemical properties of monoclonal antibodies (mAbs) is a critical process to ensure their quality, efficacy, and safety. For this purpose, mAb analysis at different levels (bottom-up, middle-up) is a common approach that includes rather complex multistep sample preparation (reduction, digestion). To ensure high analysis performance, the development of fully integrated methodologies is highly valuable.

Fast in-line bottom-up analysis of monoclonal antibodies: Toward an electrophoretic fingerprinting approach.

For their characterization and quality control, monoclonal antibodies are frequently analyzed at the bottom-up level to generate specific fingerprints that can be used to tackle post-translational modifications or ensure production consistency between lots. To circumvent time-consuming and labor-intensive off-line sample preparation steps, the implementation of integrated methodologies from sample preparation to separation and detection is highly valuable. In this perspective, capillary zone electrophoresis appears as a choice technique since the capillary can subsequently be used as a vessel for sample preparation and electrophoretic discrimination/detection of the reaction products.

A fully automated on-line salting-out assisted liquid-liquid extraction capillary electrophoresis methodology: Application to tyrosine kinase inhibitors in human plasma.

A fully automated analytical methodology combining salting-out assisted liquid-liquid extraction (SALLE) and capillary electrophoresis (CE) for the analysis of three Tyrosine Kinase Inhibitors (TKIs) in plasma samples is proposed. The automated methodology, called A-SALLE-CE-UV, makes full use of the advantages of both techniques by combining desalting, protein precipitation, automated liquid-liquid extraction, in-line CE stacking and electrophoretic separation of analytes in plasma samples in a fully integrated way. At first, the capillary is used to deliver appropriate micro-volumes of extraction agent solutions (acetonitrile, salt) in the plasma sample.

In-line tryptic digestion of therapeutic molecules by capillary electrophoresis with temperature control.

The main purpose of the present work is to provide a fully integrated temperature control of in-line tryptic digestion in order to facilitate the quality control of polypeptidic therapeutic compounds. The in-line enzymatic reaction was performed in 100 mM bicarbonate ammonium whereas a mixture of citric acid/ε-amino caproic acid (pH 5.0 and I 75 mM) was used as a background electrolyte (BGE).

Coupling of salting-out assisted liquid-liquid extraction with on-line stacking for the analysis of tyrosine kinase inhibitors in human plasma by capillary zone electrophoresis.

Developing an easy to use, cheap and fast analytical methodology is highly demanded for clinical practices, such as therapeutic drug monitoring (TDM). The present work deals with the development of an analytical methodology for the analysis of four basic anticancer drugs, namely tyrosine kinase inhibitors (TKIs), in human plasma by combining salting-out assisted liquid-liquid extraction (SALLE) with capillary electrophoresis (CE). This SALLE-CE methodology makes a full use of the advantages of both techniques by combining extraction, on-line concentration and separation in a simple way.

Integrated microreactor for enzymatic reaction automation: An easy step toward the quality control of monoclonal antibodies.

The main purpose of the present work is to provide a fully integrated miniaturized electrophoretic methodology in order to facilitate the quality control of monoclonal antibodies (mAbs). This methodology called D-PES, which stands for Diffusion-mediated Proteolysis combined with an Electrophoretic Separation, permits to perform subsequently mAb tryptic digestion and electrophoresis separation of proteolysis products in an automated manner. Tryptic digestion conditions were optimized regarding the influence of enzyme concentration and incubation time in order to achieve similar enzymatic digestion efficiency to that obtained with the classical methodology (off-line).

Evaluation of capillary zone electrophoresis for the quality control of complex biologic samples: Application to snake venoms.

Snake venoms constitute a very promising resource for the development of new medicines. They are mainly composed of very complex peptide and protein mixtures, which composition may vary significantly from batch to batch. This latter consideration is a challenge for routine quality control (QC) in the pharmaceutical industry.

Determination of artemether and lumefantrine in anti-malarial fixed-dose combination tablets by microemulsion electrokinetic chromatography with short-end injection procedure.

Background: Artemether-lumefantrine (AL) combination therapy is now the most used anti-malarial treatment in the world. Quality control of AL formulations is still a major challenge in developing countries. Until now, only liquid chromatographic methods have been reported in the literature for their analysis.

Capillary electrophoresis for the assay of fixed-dose combination tablets of artesunate and amodiaquine.

Background: Quality control of drugs in formulations is still a major challenge in developing countries. For the quality control of artesunate and amodiaquine tablets in fixed-dose combination, only liquid chromatographic methods have been proposed in the literature. There are no capillary electrophoretic methods reported for the determination of these active substances, although this technique presents several advantages over liquid chromatography (long lifetime, low price of the capillary, low volumes of electrolyte consumption) in addition to simplicity.