High‐speed separation of proteins by sodium dodecyl sulfate‐capillary gel electrophoresis with partial translational spontaneous sample injection

Abstract

In this study, we developed a picoliter-scale partial translational spontaneous injection approach which is suitable for high-speed protein separation under sodium dodecyl sulfate-capillary gel electrophoresis mode. On the basis of this approach, we built a high-speed CE system for protein separation based on a short capillary and slotted-vial array. The system has the advantages of simple structure, ease of building without the requirement of microfabricated devices, convenient operation, and low cost. Under the optimized conditions, picoliter-scale sample plugs (corresponding to ∼65 μm plug length) were obtained, which ensured both the high speed and the high efficiency in protein separation. Five fluorescein isothiocyanate labeled proteins including myoglobin, egg albumin, bovine serum albumin, phosphorylase b, and myosin were separated within 60 s with an effective separation length of 1.5 cm. Theoretical plates per meter ranging from 2.58×10⁵ to 1.28×10⁶ (corresponding to 0.78-3.88 μm plate height) were obtained. The separation speed and separation efficiency of the present system are comparable to those of most microchip-based capillary electrophoresis systems for protein separation. The relative standard deviations of the migration times were in the range of 0.9-1.3% (n=5). Good linear relationships between log relative molecular mass and migration time were obtained in the molecular weigh range of 17,200-500,000, which demonstrate the present system can be applied in protein relative molecular mass determination.

Capillary Electrophoresis in Nanotechnologies versus Nanotechnologies in Capillary Electrophoresis

“…Nowadays, an increasing number of researchers perform CE separations in short capillaries (units of centimeters) instead of microfluidic chips [48,49]. In such capillaries, fast and efficient separations are carried out without complicated chip preparation requiring expensive facilities (e.g.…”

Study of the effect of culture mediums on the amino acid metabolites for Corynebacterium glutamicum using high‐speed micellar electrokinetic chromatography

“…These advantages make CE very suitable for the analysis of AAs . Developed in recent years, high‐speed CE (HSCE), including MEKC mode, was a quick separation technique with ultrahigh separation efficiency and lesser sample consumption . Having amino groups, AAs could be derived by FITC and then detected by LIF detector .…”

Separation and determination of peptide metabolite of Bacillus licheniformis in a microbial fuel cell by high-speed capillary micellar electrokinetic chromatography

“…Fang et al. proposed a picoliter‐scale spontaneous sample injection technique for HSCE and obtained ultra‐high column efficiency in a very short separation time . In our previous work , a manual sample introduction device of high‐speed capillary micellar electrokinetic chromatography (HSCMEKC) has been introduced to analyze AAs in laver.…”

“…These advantages make CE very suitable for analyzing these compounds including AAs and peptides [15][16][17][18][19][20][21][22][23][24][25][26][27]. Fang et al proposed a picoliterscale spontaneous sample injection technique for HSCE and obtained ultra-high column efficiency in a very short separation time [28,29]. In our previous work [30], a manual sample introduction device of high-speed capillary micellar electrokinetic chromatography (HSCMEKC) has been introduced to analyze AAs in laver.…”