Separation of monoclonal antibody charge state variants by open tubular capillary electrochromatography with immobilised protein as stationary phase

Recent developments in open tubular capillary electrochromatography from 2016 to 2017

“…Bovine serum albumin (BSA) has been a well‐studied biomaterial due to its enantioselective abilities. The group of Jia fabricated a BSA‐coated poly(diallydimethylammonium chloride) (PDDA) column to investigate the selectivity of BSA in separating the charge variants of monoclonal antibodies (mAbs) for their heterogeneity characterisation . The same group conducted further improvements in simplifying column preparations by adopting fibrin as a novel SP for characterisation of the same mAbs charge variants .…”

“…Exploiting the charged state of ionised polymers or polyelectrolytes to adhere to a pre‐treated inner capillary wall has also been employed for the fabrication of SPs from nanoparticles and biomaterials . Poly(diallydimethylammonium chloride) (PDDA) was used by Zhang and co‐workers to support the fabrication of a BSA‐coated OT column by electrostatic self‐assembly . This was achieved by sequential flushing of PDDA solution in Tris‐HCl buffer (pH 8.3) containing 0.2 M NaCl, followed by BSA solution in Tris‐HCl buffer (pH 7.4).…”

Determination of protein charge variants with (imaged) capillary isoelectric focusing and capillary zone electrophoresis

“…CEC is a hybrid technique between CE and HPLC and was recently used by Zhang et al. for the separation of charge variants . There are also multidimensional approaches, for example CE‐MS , HPLC‐MS , or CE‐CE .…”

Applications of polydopamine modifications in capillary electrophoretic analysis

“…Hysteresis‐induced fluctuations in EOF and positively charged analytes adsorption are two main factors affecting the applications of CE when using bare fused silica capillary as separation column . Therefore, various surface modification strategies have been introduced in CE to extend its practical uses, including chemical coating , dynamic coating , and physical coating . Among these approaches, physical coating is a widely used method for surface modification, such as coating with positively charged polymer , hydrophilic carboxymethyl chitosan , and nanosilica .…”

“…Hysteresis-induced fluctuations in EOF and positively charged analytes adsorption are two main factors affecting the applications of CE when using bare fused silica capillary as separation column [4]. Therefore, various surface modification strategies have been introduced in CE to extend its practical uses, including chemical Article Related Abbreviations: APS, ammonium persulfate; ATRP, atom transfer radical polymerization; COF, covalent-organic framework; DA, Dopamine; DHI, 5,6-dihydroxyindole; DNA, deoxyribonucleic acid; GO, graphene oxide; LBL, layer-by-layer; LDH, layered double hydroxide; MIP, molecularly imprinted polymer; MNP, magnetic nanoparticle; NP, nanoparticle; OT-CEC, open-tubular capillary electrochromatography; OVA, ovalbumin; PAM, poly(acrylamide); PDA, polydopamine; PDMS, polydimethylsiloxane; PEEK, poly(ether ether ketone); PEI, polyetheylenimine; PMOXA, poly(2-methyl-2-oxazoline); PNE, poly (norepinephrine); rhEPO, recombinant human erythropoietin; SPR, surface plasma resonance; ZIF, zeolite imidazolate framework; β-CD, β-cyclodextrin coating [5], dynamic coating [6], and physical coating [7]. Among these approaches, physical coating is a widely used method for surface modification, such as coating with positively charged polymer [8], hydrophilic carboxymethyl chitosan [9], and nanosilica [10].…”