Simultaneous detection of assembly and disassembly of multivalent HA tag and anti‐HA antibody in single in‐capillary assay

Wang, Jianhao; Qin, Yuqin; Qin, Haifang; Liu, Li; Ding, Shumin; Teng, Yiwan; Ji, Junling; Qiu, Lin; Jiang, Pengju

doi:10.1002/elps.201600004

Cited by 7 publications

(4 citation statements)

References 31 publications

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“…The same instrumentation, methodology, and BGE were employed for investigation of a large series of interactions (self‐assembly) between oligo‐ and polypeptides of different lengths and charges (mostly tagged with hexahistidine anchor) and CdSe/ZnS quantum dots or between peptides conjugated to these quantum dots and other peptides or proteins, e.g. characterization of self assembly of glutathione stabilized CdSe/ZnS quantum dots and cyanine5‐labeled peptide (cyanine5‐Glu(Ala) 5 (His) 6 ) , detection of assembly and imidazole‐induced disassembly of multivalent HA tag peptide with hexahistidine at C‐terminus and anti‐HA monoclonal antibody , and study of multivalent interactions between conjugate of quantum dots with c‐Myc peptide tag and the anti‐c‐Myc monoclonal antibody . In the last two cases, the assays provided also the dissociation constants and stoichiometry of the complexes.…”

Section: Applicationsmentioning

confidence: 99%

Recent developments in capillary and microchip electroseparations of peptides (2015–mid 2017)

Kašička

2017

Electrophoresis

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The review brings a comprehensive overview of recent developments and applications of high performance capillary and microchip electroseparation methods (zone electrophoresis, isotachophoresis, isoelectric focusing, affinity electrophoresis, electrokinetic chromatography, and electrochromatography) to analysis, microscale isolation, purification, and physicochemical and biochemical characterization of peptides in the years 2015, 2016, and ca. up to the middle of 2017. Advances in the investigation of electromigration properties of peptides and in the methodology of their analysis (sample preseparation, preconcentration and derivatization, adsorption suppression and EOF control, and detection) are described. New developments in particular CE and CEC methods are presented and several types of their applications to peptide analysis are reported: qualitative and quantitative analysis, determination in complex (bio)matrices, monitoring of chemical and enzymatical reactions and physical changes, amino acid, sequence and chiral analysis, and peptide mapping of proteins. Some micropreparative peptide separations are shown and capabilities of CE and CEC methods to provide important physicochemical characteristics of peptides are demonstrated.

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Section: Applicationsmentioning

confidence: 99%

Recent developments in capillary and microchip electroseparations of peptides (2015–mid 2017)

Kašička

2017

Electrophoresis

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“…However, the direct effect of the analyte on the fluorescent properties of QDs in combination with changes in the electrophoretic mobility can also provide valuable information on their mutual interactions as well as the efficiency of the bioconjugation or self‐assembly characterization of QDs .…”

Section: Ce Separations Of Nanomaterialsmentioning

confidence: 99%

Capillary electrophoresis and nanomaterials – Part I: Capillary electrophoresis of nanomaterials

Adam

Vaculovičová

2017

Electrophoresis

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Nanomaterials are in analytical science used for a broad range of purposes, covering the area of sample pretreatment as well as separation, detection, and identification of target molecules. This part of the review covers capillary electrophoresis (CE) of nanomaterials and focuses on the application of CE as a method for characterization used during nanomaterial synthesis and modification as well as the monitoring of their properties and interactions with other molecules. The heterogeneity of the nanomaterial family is extremely large. Depending on different definitions of the term Nanomaterial/Nanoparticle, the group may cover metal and polymeric nanoparticles, carbon nanomaterials, liposomes and even dendrimers. Moreover, these nanomaterials are usually subjected to some kind of surface modification or functionalization, which broadens the diversity even more. Not only for purposes of verification of nanomaterial synthesis and batch-to-batch quality check, but also for determination the polydispersity and for functionality characterization on the nanoparticle surface, has CE offered very beneficial capabilities. Finally, the monitoring of interactions between nanomaterials and other (bio)molecules is easily performed by some kind of capillary electromigration technique.

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“…Expression of eGFP did not prevent cell division and was persistent in cell division, like HEV [9]. Since its introduction almost 20 years ago as a short immunoreactive tag, the hemagglutinin epitope YPYDVPDYA (HA tag) from influenza virus A has been used to track proteins of interest within cells, to isolate them, and to coprecipitate their possible partners [10,11]. Using the HA-tag replicons, HA-tag protein cloned into aHEV was expressed in LMH cells.…”

mentioning

confidence: 99%

Construction of an avian hepatitis E virus replicon expressing heterologous genes and evaluation of its potential as an RNA vaccine platform

et al. 2021

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Avian hepatitis E virus (aHEV) has a 5′-capped, positive-sense, single-stranded RNA genome of approximately 6.6 kb and consists of 5′-non-coding regions (NCR), three open reading frames (ORFs), and a 3′ NCR. ORF1 encodes the non-structural proteins, ORF2 encodes an immunogenic capsid protein, and ORF3 encodes a phosphoprotein [1,2]. Infectious cDNA clones that can synthesize the original viral RNA are useful for virus characterization and pathogenicity research. They are also useful vaccine platforms; a replicon can be created by inserting a gene of interest into the structural gene [3]. The RNA genomes of positive-strand RNA viruses have dual functions: they act as an mRNA template for RNA-dependent RNA polymerase (RDRP) and as a genomic template for replication by the respective RDRP. Replicon RNA is readily produced by in vitro transcription from cDNA templates [4,5]. However, no study has reported a replicon vaccine platform using aHEV. Therefore, we explored the possibility of aHEV as a vaccine platform using an aHEV infectious cDNA clone. The ORF2 gene of aHEV was replaced by heterologous genes, such as enhanced green fluorescent protein (eGFP), hemagglutinin (HA)-tag, and infectious bronchitis virus (IBV) S1. The S1 protein of IBV is critical for antigenic neutralization and hemagglutination. IBV infection is a significant problem in the poultry industry, and novel vaccines are needed to combat emerging and variant IBV serotypes [6]. These replicons were constructed and used to transcribe the capped RNA transcripts and to transfect leghorn male hepatoma (LMH) cells. This is the first study of a replicon vaccine platform using aHEV infectious cDNA clones.The previously constructed genotype 2 aHEV infectious cDNA pT9-aHEV-GI clone and the genotype 1 aHEV infectious cDNA pT11-aHEV-K clone were used in this study [7]. Specific primers were prepared to insert eGFP and an HA-tag into the pT9-aHEV-GI clone (Table 1). The products were subjected to fusion

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Simultaneous detection of assembly and disassembly of multivalent HA tag and anti‐HA antibody in single in‐capillary assay

Cited by 7 publications

References 31 publications

Recent developments in capillary and microchip electroseparations of peptides (2015–mid 2017)

Recent developments in capillary and microchip electroseparations of peptides (2015–mid 2017)

Capillary electrophoresis and nanomaterials – Part I: Capillary electrophoresis of nanomaterials

Construction of an avian hepatitis E virus replicon expressing heterologous genes and evaluation of its potential as an RNA vaccine platform

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