Improving the capillary electrophoretic analysis of poliovirus using a Plackett–Burman design

Oita, Iuliana; Halewyck, Hadewych; Pieters, Sigrid; Dejaegher, Bieke; Thys, Bert; Rombaut, Bartholomeus; Heyden, Yvan Vander

doi:10.1016/j.jpba.2008.09.049

Cited by 21 publications

(35 citation statements)

References 38 publications

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“…Poliovirions collected after sucrose gradient ultracentrifugation were analyzed with CE using the method described in [16]. The electropherogram is shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Dimethylformamide was added (0.0001%) as EOF marker in the nonsucrosepurified samples. In the sucrose matrix samples, the sucrose itself migrates with the EOF, generating a negative peak, and therefore no EOF marker was added anymore [16].…”

Section: Pvmentioning

confidence: 99%

“…First attempts to analyze PV with CE were performed with a small sample plug injection using separation conditions selected from the literature which were further optimized [16]. In short, PV was successfully analyzed by CE in a moderate alkaline 50 mM borate buffer of pH 8.3 as BGE.…”

Section: Introductionmentioning

confidence: 99%

“…Therefore, different detergents in the BGE were tested from which SDS seemed to be the most successful to avoid analyte-wall interactions and to achieve reproducible results. After the application of a Plackett-Burman design, a good resolution between the PV peak and the system peaks was achieved [16]. The drawback of SDS is, however, a possible destruction of the investigated biomolecules, which can be seen as protein assemblies.…”

Section: Introductionmentioning

confidence: 99%

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Identification of poliovirions and subviral particles by capillary electrophoresis

et al. 2010

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Poliovirions, purified from infected cell extracts with anion-exchange chromatography, can be analyzed and identified by CE in untreated fused silica capillaries using UV detection. Other subviral particles can be eluted as well from the same infected cell extract using a higher salt concentration buffer on the ion-exchange chromatography. Virions can be identified because of their conversion into empty capsids upon heating at 561C. As a result of heating, the viral genome is released from the capsid. Here, we show that during this incubation some intermediate particles were found. The latter were identified by enzymatic peak shift analysis. The high salt concentration eluate subviral particles were analyzed with preincubation affinity CE together with their sensitivity for RNase and proteinase K treatment. Electropherograms of the higher salt concentration eluate display a mixture of at least four different subviral particles. One particle proved to have an [N1, H] antigenicity and was resistant to RNase and proteinase K digestion. The remaining particles were all sensitive to proteinase K treatment. This CE method proved to be valuable in the detection, identification and analysis of poliovirions and poliovirus particles offering an alternative powerful, cheap, fast and easy analysis method.

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“…Poliovirions collected after sucrose gradient ultracentrifugation were analyzed with CE using the method described in [16]. The electropherogram is shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

Section: Pvmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Identification of poliovirions and subviral particles by capillary electrophoresis

et al. 2010

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“…Thus, Oita et al [119] developed a CE method combined with 12-experiment Plackett-Burman design for analyzing poliovirus. They reduced the detection concentration of poliovirus to 140 μg/mL.…”

Section: Detecting and Diagnosing Virusesmentioning

confidence: 99%

Capillary electrophoresis based on nucleic acid detection for diagnosing human infectious disease

Lian

Zhao

2016

Clinical Chemistry and Laboratory Medicine (CCLM)

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Rapid transmission, high morbidity, and mortality are the features of human infectious diseases caused by microorganisms, such as bacteria, fungi, and viruses. These diseases may lead within a short period of time to great personal and property losses, especially in regions where sanitation is poor. Thus, rapid diagnoses are vital for the prevention and therapeutic intervention of human infectious diseases. Several conventional methods are often used to diagnose infectious diseases, e.g. methods based on cultures or morphology, or biochemical tests based on metabonomics. Although traditional methods are considered gold standards and are used most frequently, they are laborious, time consuming, and tedious and cannot meet the demand for rapid diagnoses. Disease diagnosis using capillary electrophoresis methods has the advantages of high efficiency, high throughput, and high speed, and coupled with the different nucleic acid detection strategies overcomes the drawbacks of traditional identification methods, precluding many types of false positive and negative results. Therefore, this review focuses on the application of capillary electrophoresis based on nucleic detection to the diagnosis of human infectious diseases, and offers an introduction to the limitations, advantages, and future developments of this approach.

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Virus analysis using electromigration techniques

2009

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We discuss the progress during the last 4 years in the analysis of viruses by electrophoresis in capillaries and microfluidic devices. The paper is the continuation of a review published in this journal in 2005 [Kremser, L., Blaas, D., Kenndler, E., Electrophoresis 2004, 25, 2282-2291]. Eighteen papers on the topic have appeared since; the majority deals with zone electrophoresis and three reports are on IEF. These methods have been applied to human rhinoviruses, poliovirus Semliki Forest virus, norovirus-like particles, and the two bacteriophages MS2 and T5. A main finding was that addition of detergents and salts to the BGEs are essential for the robustness of the CE analysis. Analyte detection was usually via UV absorbance but there are some examples where the viruses were rendered fluorescent via modification of the capsid proteins with reactive dyes and/or by non-covalent attachment of intercalating fluorescent compounds to the nucleic acids making up the viral genome. Interestingly, some viruses are permeable to small molecular mass components; this allows fluorescent dyes to diffuse into the intact virus where they attach to the nucleic acid. Release of a viral genome upon heating was also monitored by using similar methodologies. Interactions of viruses and subviral particles with antibodies, receptors, and receptor-decorated liposomes were investigated with CE methods, all by using a non-equilibrium approach (i.e. co-incubation of the components prior to CE separation). Viruses are multivalent (i.e. possess many identical surface-exposed patches) and most of them are composed of defined numbers of identical subunits. The high resolution of CE has been most remarkably demonstrated by the separation of stoichiometric complexes between virus and a distinct number of soluble recombinant receptors and revealed their concentration-dependent distribution.

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Improving the capillary electrophoretic analysis of poliovirus using a Plackett–Burman design

Cited by 21 publications

References 38 publications

Identification of poliovirions and subviral particles by capillary electrophoresis

Identification of poliovirions and subviral particles by capillary electrophoresis

Capillary electrophoresis based on nucleic acid detection for diagnosing human infectious disease

Virus analysis using electromigration techniques

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