Capillary isoelectric focusing of proteins and microorganisms in dynamically modified fused silica with UV detection

This review article with 239 references describes recent developments in capillary electrophoresis of proteins, and covers the two years since the previous review (V. Dolník, Electrophoresis 2006, 27, 126-141) through spring 2007. It includes topics related to CE of proteins, such as sample pretreatment, wall coatings, improving separation, various forms of detection, and special electrophoretic techniques including ACE, CIEF, capillary ITP, and CEC. The paper describes applications of CE to analysis of proteins in real-world samples including human body fluids, food and agricultural samples, protein pharmaceuticals and recombinant protein preparations.

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“…CIEF of proteins and microorganisms has been performed in fused-silica capillaries with a dynamic PEG coating and UV detection [70].…”

Section: Capillary Iefmentioning

confidence: 99%

Capillary electrophoresis of proteins 2005–2007

Dolnı́k¹

2007

Electrophoresis

152

106

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“…However, the detection of protein contamination from AF in individual fractions of sucrose gradient and in the sucrose cushion, after precipitation of viruses in AF by means of PEG, was significantly different. Generally, after ultracentrifugation [45,48], the majority of protein contamination occurs in the fractions with the lowest density. After PEG precipitation, the greater part of contamination was present in the fractions with the highest, 40% w/v, and lowest density of sucrose, (15% w/v in the case of EQUI, see Fig.…”

Section: Comparison Of the Results From Cief And Qrt-pcr Techniques Omentioning

confidence: 99%

“…The sample injection was accomplished by siphoning action obtained by elevating of the inlet reservoir on the side of the anode relative to the outlet reservoir on the side of the cathode. The segmental injection of the sample into the capillary [45] was employed here. The height difference of the reservoirs for the sample injection, Wh, was 100 mm, and the time of the injection, t inj.…”

Section: Equipment and Proceduresmentioning

confidence: 99%

“…solution of the selected simple ampholytic electrolytes dissolved in the catholyte [45,46], HEPES (pK 1 5 3.65, pK 2 5 7.5, pI 5 5.58) and L-aspartic acid (pK 1 5 1.88, pK 2 5 3.66, pI 5 2.77) in the ratio 1:1, the segment of the sample mixture of viruses, and the segment of both the mixture of commercial carrier ampholytes and the UV detectable pI markers used for the tracing of the pH gradient in the range 2.0-7.5 or 3.0-10.0. The time of the injection of the spacer segment was 25 s, the sample segment 5-15 s and the segment of carrier ampholytes and pI markers was 40 s.…”

Section: Sample Preparationmentioning

confidence: 99%

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Testing of the influenza virus purification by CIEF

et al. 2010

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In virological practice, the pre-concentration, purification and subsequent determination of the purity and concentration of the viruses from the cultural medium and/or from the real sample are required. The conventional techniques used today are equipment demanding, time-consuming and laborious. In this study, the CIEF of influenza viruses with UV detection has been developed and subsequently used to test the purification of the virus from the biological samples. The equine and swine influenza viruses present in infected allantoic fluid of specific pathogen free embryonated chicken eggs were precipitated by using PEG 6000 and sodium chloride. The precipitated viruses were centrifuged at 14 000 x g, and the impurities of different densities were removed by using the sucrose gradients. The efficiency of the virus purification technique was examined by the CIEF and compared to the results of real-time PCR. The pIs of both influenza viruses were determined. Simultaneously, the CIEF was found to be a suitable method for the rapid testing of the efficiency of the virus purification.

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“…polyethylene oxide [22][23][24][25] or PEG [26]) with only one report of using capillaries with covalent modifications using trimethylchlorosilane and divinylbenzene [22]. One particular dynamic coating based on PEG 4000 was used for CIEF of microorganisms that were UV detected after focusing [26].…”

Section: Microorganismsmentioning

confidence: 99%