Protein A immobilized monolithic capillary column for affinity chromatography

Pan, Zaifa; Zou, Hanfa; Mo, Weimin; Huang, Xiaodong; Wu, Ren’an

doi:10.1016/s0003-2670(02)00511-1

Cited by 83 publications

(70 citation statements)

References 30 publications

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“…The majority of the reported monolithic columns for separation and analysis of glycans are organic polymer-based [82][83][84][85][86][87]89,90,[92][93][94][95][97][98][99][100][101][102][103][104][107][108][109][110][111][112]114,115,[118][119][120][121]123,[144][145][146]. This may be due to the straightforward approach in the preparation and the availability of a variety of functional monomers to obtain desired functionality of the surface.…”

Section: Monolithic Columns Can Easily Be Prepared In-situmentioning

confidence: 99%

“…An example of post-modification was photografting of hydrophilic polymer poly(ethyleneglycol) methacrylate (PEGMA) to reduce the hydrophobicity of the monolith [82]. To create affinity mode, lectins were immobilized on the surface in various ways such as covalent binding of amine residues to aldehydefunctionalized surfaces [84,98,115,117,[119][120][121], chelation to (IDA-Cu 2+ ) modified surfaces [99] and a molecular imprinting method using a polydopamine coating [112]. Coating a monolith surface with nanoparticles provides a high loading of functional groups, fixing the problem of limited active sites described above.…”

Section: Monolithic Columns Are Versatile To a Variety Of Available Smentioning

confidence: 99%

“…The advantage of this method over conventional SELEX is that it is economical requiring only 10-20 µL of target and library DNA per cycle [114]. A monolithic capillary column with covalently immobilized protein A was used to determine the hIgG concentration in human serum [115]. Protein A is an expensive protein.…”

Section: Boronate Affinitymentioning

confidence: 99%

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Development of Monolithic Column Materials for the Separation and Analysis of Glycans

Alla

Stine

2015

Chromatography

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Monolithic column materials offer great advantages as chromatographic media in bioseparations and as solid-supports in biocatalysis. These single-piece porous materials have an interconnected ligament structure that limits the void volume inside the column, thus increasing the efficiency without sacrificing the permeability. The preparation of monolithic materials is easy, reproducible and has available a wide range of chemistries to utilize. Complex, heterogeneous and isobaric glycan structures require preparation methods that may include glycan release, separation and enrichment prior to a comprehensive and site-specific glycosylation analysis. Monolithic column materials aid that demand, as shown by the results reported by the research works presented in this review. These works include selective capture of glycans and glycoproteins via their interactions with lectins, boronic acids, hydrophobic, and hydrophilic/polar functional groups on monolith surfaces. It also includes immobilization of enzymes trypsin and PNGase F on monoliths to digest and deglycosylate glycoproteins and glycopeptides, respectively. The use of monolithic capillary columns for glycan separations through nano-liquid chromatography (nano-LC) and capillary electrochromatography (CEC) and coupling these columns to MS instruments to create multidimensional systems show the potential in the development of miniaturized, high-throughput and automated systems of glycan separation and analysis.

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Section: Monolithic Columns Can Easily Be Prepared In-situmentioning

confidence: 99%

Section: Monolithic Columns Are Versatile To a Variety Of Available Smentioning

confidence: 99%

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Development of Monolithic Column Materials for the Separation and Analysis of Glycans

Alla

Stine

2015

Chromatography

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“…The use of other monoliths for the separation of proteins and peptides in capillary chromatography is documented in a number of publications. Mayr et al [30] separated in only 5 min a mixture of six proteins on norbornene-based capillary monoliths and Pan et al [31] described the purification of human immunoglobulin G (IgG) affinity chromatography in capillary columns of glycidyl methacrylate. As we could recently show, glycidyl methacrylate monoliths can be used for reversed-phase chromatography of proteins with high separation efficiency (Fig.…”

Section: Capillary Chromatographymentioning

confidence: 99%

Recent progress in high‐performance capillary bioseparations

et al. 2003

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Bioanalysis is a fast growing domain with new technological developments in the field of mass spectrometry, separation science, and bioinformatics. The central role of separation prior to detection and evaluation of analytes is indisputable. In fact, since the decipherment of the human genome via multichannel capillary electrophoresis in combination with laser-induced fluorescence [1], the analysis of the proteome, i.e., the type and amount of all proteins expressed at a certain time by the genome in a cell or tissue [2] became of central interest. Therefore, new fast and sensitive capillary liquid chromatographic, electrophoretic, and electrochromatographic separation techniques, suitable for high-throughput analysis of proteins, peptides and nucleic acids, are required.

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“…Examples include the immobilisation of protein A, for the separation of immunoglobulins [6][7][8], lectins such as concavalin A, for the extraction of glycoproteins [9], enzymes such as trypsin, for use as microbioreactors in LC-MS proteomic applications [4], and the immobilisation of immunoglobulins [10]. These varied applications of polymeric monolithic stationary phases for affinity chromatography has been the subject of a recent review by Mallik [11] in which the numerous immobilisation strategies which have previously been reported are discussed.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of photografted charged sites within polymer monoliths in capillary columns using contactless conductivity detection

Connolly

O'Shea

Clark

et al. 2007

J of Separation Science

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Original PaperEvaluation of photografted charged sites within polymer monoliths in capillary columns using contactless conductivity detection Capacitively coupled contactless conductivity detection (C 4 D) is presented as a novel and versatile means of visualising discrete zones of charged functional groups grafted onto polymer based monoliths. Monoliths were first formed within 100 lm UV-transparent fused silica capillaries. Photografting methods were subsequently used to graft a charged functional monomer, 2-acrylamido-2-methyl-1-propanesulfonic acid, onto discrete regions of the monolith using a photomask. Post-modification monolith evaluation involves scanning the C 4 D detector along the length of the monolith to obtain a profile of the exact spatial location of grafted charged functionalities with millimetre accuracy. The methodology was extended to the visualisation of several zones of immobilised protein (bovine serum albumin) using photografted azlactone groups to enable covalent attachment of the protein to the monolith at precise locations along its length. In addition, the extent of non-specific binding of protein to the ungrafted regions of the monolith due to hydrophobic interactions could be monitored as an increase in background conductivity of the stationary phase. Finally, the technique was cross-validated using digital photography in combination with a UV light source by immobilising green fluorescent protein in discrete zones and comparing the results obtained using both complementary techniques.

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Protein A immobilized monolithic capillary column for affinity chromatography

Cited by 83 publications

References 30 publications

Development of Monolithic Column Materials for the Separation and Analysis of Glycans

Development of Monolithic Column Materials for the Separation and Analysis of Glycans

Recent progress in high‐performance capillary bioseparations

Evaluation of photografted charged sites within polymer monoliths in capillary columns using contactless conductivity detection

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