High‐throughput capillary electrophoresis analysis of biopharmaceuticals utilizing sequential injections

Abstract

The complexity of biotherapeutic products implies an ever‐increasing list of product quality attributes that need to be monitored and characterized. In addition, the growing interest in implementing process analytical technology in biopharmaceutical production has further increased the testing burden, together with the need for rapid testing that can facilitate real‐time or near‐real‐time decision‐making. Capillary electrophoresis (CE) has made a place in biopharmaceutical analysis but is regarded as a low‐throughput method, with the instrument dead time constituting more than 80% of the total time of analysis. In this study, the dead time of CE was utilized to analyse 3 mAb samples in a single‐CE run. This approach resulted in an up to 77% reduction in the total analysis time and increased the productivity by up to 300%, compared to traditional single CE‐ultraviolet runs, without compromising resolution or relative peak areas. Additionally, good method reproducibility was observed. The compatibility of the method has been demonstrated with protein A eluate and cation exchange chromatography fractions. We, thus, propose that sequential injections can be applied for fast and robust CE analysis of biopharmaceuticals.

Recent developments in capillary and microchip electroseparations of peptides (2021–mid‐2023)

“…In this mode, the analyses of the preceding sample are stopped after some time (separation voltage is switched off), a new sample is introduced, separation voltage is switched on, and the analysis of both samples continues. Using this approach, the analyses of peptide in pharmaceutical and biological matrices [91] and qualitative analysis of biopharmaceuticals [132] were increased three times, and the time analysis of one sample was reduced the same time.…”

Identity verification of monoclonal antibodies by triple injection capillary zone electrophoresis

“…In addition, early migrating peaks originating from components in the pharmaceutical formulation are used to confirm the product similarity, which might be helpful to distinguish genuine mAb drugs from suspect counterfeits. Only one study with multiple‐injection have analyzed mAb physiochemical characteristics with UV detection, but limited their scope to analysis time, resolution, relative peak areas and reproducibility [14].…”

“…First, permanently or dynamically, coated capillaries can be used [10,11]. Second, multiple injection, where several samples are analyzed at the same conditions in a single run, can eliminate inter-run variations in migration times [12,13] and decrease total run time [14]. Multiple injection in CE, also denoted as multisegment injection or multiple sequential injection, separation window-dependent multiple injection and repeated sample injection have previously been used for analysis of small molecules [15][16][17][18][19][20][21][22][23][24], proteins [25,26], mAbantigen interaction with UV [27], or mAb glycosylation with laser-induced fluorescence detector [28].…”

Numerical modeling reveals improved organelle separation for dielectrophoretic ratchet migration

“…However, there are currently only limited approaches for fractionating organelles with high separation efficiency and throughput. The few size-based separation techniques reported for mitochondria include free-flow fractionation [10] and capillary electrophoresis [11][12][13]. Dielectrophoresis (DEP) is another widely studied particle separation technique used for the fractionation of sub-micrometer-sized particles.…”