A novel core-shell type polymer support for solid-phase peptide synthesis

Cho, Jin Ku; Park, Byeong-Deog; Lee, Yoon‐Sik

doi:10.1016/s0040-4039(00)01279-x

Cited by 24 publications

(15 citation statements)

References 14 publications

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“…As previously reported, the CutiCore bead showed a fast reaction rate in the photo-cleavage reactions because most of the functional groups are located at the shell of the bead [37]. The photo-cleavable ligand-coupled CutiCore beads were packed into a microfilter chamber (volume: 0.4 mL) within a microchip (Fig.…”

Section: Resultsmentioning

confidence: 97%

Microaffinity purification of proteins based on photolytic elution: Toward an efficient microbead affinity chromatography on a chip

Chung¹,

Kim²,

Cho

et al. 2005

Electrophoresis

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A bead affinity chromatography system, which was based on the photolytic elution method, was integrated into a glass-silicon microchip to purify specific target proteins. CutiCore beads, which were coupled with a photo-cleavable ligand, such as biotin and an RNA aptamer, were introduced into a filter chamber in the microchip. The protein mixture containing target protein labeled with fluorescein isothiocyanate (FITC) was then passed through the packed affinity beads in the microchamber by pressure-driven flow. During the process, the adsorbed protein on the bead was monitored by fluorescence. The concentrated target protein on the affinity bead was released by simple irradiation with UV light at a wavelength of 360 nm, and subsequently eluted with the phosphate buffer flow. The eluted target protein was quantitatively detected via the fluorescence intensity measurements at the downstream of the capillary connected to the outlet of the microchip. The microaffinity purification allowed for a successful method for the identification of specific target proteins from a protein mixture. In addition, the feasibility of this system for use as a diagnosis chip was demonstrated.

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Section: Resultsmentioning

confidence: 97%

Microaffinity purification of proteins based on photolytic elution: Toward an efficient microbead affinity chromatography on a chip

Chung¹,

Kim²,

Cho

et al. 2005

Electrophoresis

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“…Core shell-type resins with a highly cross-linked polystyrene core and a poly(ethylene glycol) (PEG) shell [25] (0.2 mmol/g, 4 mol% XL, 200-400 mesh) were preswollen in DMF at room temperature for 1 h. 4-(4-(1-Hydroxyethyl)-2-methoxy-5-nitrophenoxy)butanoic acid (3 eq), BOP (3 eq), and DIEA (3 eq) in dry DMF were added to a suspension of the resin (1 eq), and the suspension was shaken at RT for 4 h. Subsequently, the resin was washed with DMF (265 mL), DCM (365 mL), and successively with MeOH, DCM, and MeOH (5 mL each). Hydroxy photocleavable linker-coupled resin was dried under reduced pressure.…”

Section: Preparation Of a Photocleavable Boc-n-phe-oh Coupled Resinmentioning

confidence: 99%

“…After finishing the reaction, the beads linked by covalent bonding with RNA oligonucleotides were mixed with 1 M ethanolamine to block the residual active sites, NHS for 2 h at RT, and were sequentially washed with 25 mM Tris-HCl containing 10 mM MgCl 2 for five times. After the aptamer was covalently bound to the magnetite Cuticore beads from BeadTech (Seoul, Korea) as stated above [25], the beads were guided to the center of the bead chamber by magnetic fields.…”

Section: Aptamer Labeling and Immobilization On Beadsmentioning

confidence: 99%

“…Then, the amount of HCV RNA polymerase in serums of an HCV-infected patient and a normal healthy person were compared using the same ELISA kit. Figure 6 shows the amount of HCV RNA polymerase in variable volumes of serum (10,25,50,70,85, and 100 mL) based on the calibration of pure HCV RNA polymerase in between 1 mg/mL and 10 mg/mL (r 2 = 0.96). One thing to remark here is that the false positive background level of HCV RNA polymerase in the serum is very high, so that RNA polymerase level below 0.22 6 0.02 mg in serum cannot be accurately measured.…”

Section: Analysis Of Hcv Rna Polymerase By Elisamentioning

confidence: 99%

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Microbead‐based affinity chromatography chip using RNA aptamer modified with photocleavable linker

Cho

Lee²,

Chung

et al. 2004

Electrophoresis

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A microbead-based affinity chromatography chip (micro-BACC) controlling hundreds of nanoliters of reaction volume was developed to separate and analyze hepatitis C virus (HCV) RNA polymerase protein by immobilization of an RNA aptamer on beads. A photocleavable linker was conjugated in between the beads and the aptamer to elute the bound RNA polymerase from the RNA aptamer in one step by UV irradiation, resulting in an efficient method to elute and identify the target molecule bound on RNA using a mass spectrometer. This linker showed a cleavage activity over 70% upon UV irradiation at 1050 mW/cm2 for more than 5 min. The photoelution method could prevent the target molecule from contaminations in affinity chromatography caused by elution solutions of high salt concentration, extreme pH and detergent, respectively. In this chip, sample reagents up to 800 nL could be metered quantitatively into the bead chamber using a nanoliter dispenser working, based on surface-guided flow control and pneumatic control by external air pressure on the chip. RNA polymerase eluted after UV irradiation was successfully analyzed by trypsin treatment without additional purification. As a result, using the aptamer, we could detect RNA polymerase from 800 nL hepatitis C patient serum containing 96 fmol HCV RNA polymerase. The detection limit of this system was estimated to be 9.6 fmol HCV RNA polymerase.

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“…[2] A study reported by Cho et al also shows that core-shell microspheres could be employed during solid-phase peptide synthesis. [3] In particular, streptavidin-coated particles have found many applications because of their selective and strong interaction with biotin. Many compounds can be biotinylated in one simple step and be bound by strong biotin/ streptavidin (or the proxy biotin/avidin) complex formation to the surface of streptavidin-coated spheres.…”

Section: Introductionmentioning

confidence: 99%

One Pot Synthesis of Surface PEGylated Core–Shell Microparticles by Suspension Polymerization with Surface Enrichment of Biotin/Avidin Conjugation

Ting¹,

Nguyen²,

Stenzel³

2009

Macromolecular Bioscience

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Core-shell microparticles that consist of poly(vinyl neodecanoate) (VND) crosslinked with poly(ethylene glycol dimethacrylate) (EGDMA) as the core and poly(ethylene glycol methacrylate) (PEGMA) (Mn = 360 or Mn = 526 g . mol(-1)) as the shell have been synthesized using suspension polymerization by a conventional free radical polymerization process. Interfacial tension and stability tests show that PEGMA acts as an amphiphilic macromonomer and is located on the oil/water interface of the suspension system, thus forming an outer layer during the polymerization. Kinetic studies of the monomers' conversion of VND, EGDMA, and PEGMA have been carried out using (1)H NMR spectroscopy. EGDMA and PEGMA were found to have faster reaction rates compared to VND. Moreover, scanning electron microscopy showed that the polymerization of these particles starts from the shell and finishes towards the core. Consequently, the resulting microsphere is found to have a multi-layer structure. Biotin was covalently bound to the surface by the PEGMA hydroxy groups. Conjugation of biotin with streptavidin PE (phycoerythrin) was subsequently carried out. Confocal microscopy was used to confirm the presence of fluorescing streptavidin. The amount of avidin conjugated to the microspheres was calculated by the release of a 2-(4-hydroxyphenylazo)benzoic acid/avidin complex using UV/vis spectroscopy. One avidin molecule was found to occupy 7 nm(2) on the surface of the microspheres.

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A novel core-shell type polymer support for solid-phase peptide synthesis

Cited by 24 publications

References 14 publications

Microaffinity purification of proteins based on photolytic elution: Toward an efficient microbead affinity chromatography on a chip

Microaffinity purification of proteins based on photolytic elution: Toward an efficient microbead affinity chromatography on a chip

Microbead‐based affinity chromatography chip using RNA aptamer modified with photocleavable linker

One Pot Synthesis of Surface PEGylated Core–Shell Microparticles by Suspension Polymerization with Surface Enrichment of Biotin/Avidin Conjugation

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