Sugar-Sensitive Thin Films Composed of Concanavalin A and Glycogen

Sato, Katsuhiko; Imoto, Yoko; Sugama, Jun; Seki, Shunsuke; Inoue, Hiroyuki; Odagiri, Tsuyoshi; Anzai, Jun Ichi

doi:10.2116/analsci.20.1247

Cited by 27 publications

(15 citation statements)

References 11 publications

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“…FITC labeling ratios for the FITC-protein conjugates were determined using FITC vendor's instructions where the FITC molar extinction coefficient at 495 nm was 84 000 M −1 cm −1 and the molar extinction coefficient at 280 nm was assumed to be 30% of the molar extinction coefficient at 495 nm. The protein molar extinction coefficient at 280 nm were 43 291, 40 057, 37 669 and 160 000 M −1 cm −1 for BSA, [16] lysozyme, [16] trypsin, [16] and Con A [17] respectively. FITC : protein labeling ratios for these proteins were determined to be 1 : 1, 0.35 : 1, 0.18 : 1, 2.15 : 1, respectively.…”

Section: Preparation Of Fitc-protein Conjugates and Fitc Labeling Ratiosmentioning

confidence: 98%

Protein adsorption drastically reduces surface‐enhanced Raman signal of dye molecules

Zhang

Ansar

Vangala

et al. 2009

J Raman Spectroscopy

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There is an increasing interest in developing surface enhancement Raman spectroscopy methods for intracellular biomolecule and for in vitro protein detection that involve dye or protein-dye conjugates. In this work, we have demonstrated that protein adsorption on silver nanoparticle (AgNP) can significantly attenuate the surface-enhanced Raman spectroscopy (SERS) signal of dye molecules in both protein/dye mixtures and protein/dye conjugates. SERS spectra of 12 protein/dye mixtures were acquired using 4 proteins [bovine serum albumin (BSA), lysozyme, trypsin, and concanavalin A] and three dyes [Rhodamine 6G, adenine, and fluorescein isothiocyanate (FITC)]. Besides the protein/dye mixtures, spectra were also obtained for the free dyes and four FITC-conjugated proteins. While no SERS signal was observed in protein/FITC mixtures or conjugates, a significantly reduced SERS intensity (up to 3 orders of magnitude) was observed for both R6G and adenine in their respective protein mixtures. Quantitative estimation of the number of dye molecules absorbed onto AgNP implied that the degree of R6G SERS signal reduction in the R6G/BSA sample is 2 to 3 orders of magnitude higher than what could be accounted for by the difference in the amount of the absorbed dyes. This finding has significant implications for both intracellular SERS analyses and in vitro protein detection using SERS tagging strategies that rely on Raman dyes as reporter molecules.

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Section: Preparation Of Fitc-protein Conjugates and Fitc Labeling Ratiosmentioning

confidence: 98%

Protein adsorption drastically reduces surface‐enhanced Raman signal of dye molecules

Zhang

Ansar

Vangala

et al. 2009

J Raman Spectroscopy

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“…It is noted that not only glycogen, 10,11 but also the synthetic polymer (i.e., PV-MA) can also be used for constructing the sugar-sensitive Con A assembly. This in turn suggests that the performance characteristics of the sugar-sensitive Con A assembly could be further improved by suitably modiying the chemistry of the polymer.…”

Section: Sugar-induced Disintegration Of Con A/pv-ma Filmmentioning

confidence: 99%

“…9 We have already reported on the preparation of stimuli-sensitive multilayer films that can be disintegrated by being exposed to sugars. 10,11 Stimuli-sensitive multilayer films were fabricated using concanavalin A (Con A) and glycogen. Con A is a lectin protein (molecular weight; 104000) found in Jack bean and is known to contain four identical binding sites to sugars, such as D-mannose and D-glucose (Fig.…”

Section: Introductionmentioning

confidence: 99%

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Sugar-Sensitive Thin Films Composed of Concanavalin A and Sugar-Bearing Polymers

2005

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Layered thin films composed of concanavalin A (Con A) and sugar-bearing polymers were prepared by a layer-by-layer deposition of Con A and the polymer on a solid surface. The sugar-induced disintegration was studied. Con A-polymer layered films could be successfully prepared using a maltose-bearing polymer (PV-MA), while melibiose-and glucosebearing polymers (PV-MEA and PV-G) did not afford a layered film, due to a weak affinity of PV-MEA and PV-G to Con A. The Con A/PV-MA layered film was stable in pH 7 and 8 solutions, while in a pH 6 medium the film was slightly unstable. The Con A/PV-MA film was disintegrated upon the addition of sugars in solution owing to a preferential binding of the sugars to the binding site of Con A in the film. The disintegration rate was dependent on the type of sugar and its concentration. The Con A/PV-MA film was disintegrated rapidly upon the addition of methyl α-Dmannopyranoside, while the rate was slower upon the addition of the same concentration of D-mannose, D-glucose and methyl α-D-glucopyranoside. The present system may be useful for constructing sensitive devices that can release a drug or other functional molecules in response to sugars.

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“…Both glucose and mannose groups carry a free -OH group at their C-4 and C-6 sites with a specific orientation, which can recognize the binding site of Con A subunit efficiently and form a highly specific 4:1 sugar-Con A complex [9][10][11][12]. Sugar-Con A interaction can also be used to assemble LBL films [8,[13][14][15][16][17][18][19][20][21][22][23][24][25]. For example, some glycoproteins such as glucose oxidase (GOD) and horseradish peroxidase (HRP) can be assembled with Con A into LBL films and retain their catalytic activity [8].…”

Section: Introductionmentioning

confidence: 99%

Loading of myoglobin into layer-by-layer films assembled by concanavalin A and dextran based on their biospecific recognition: An electrochemical study

Yao¹,

Guo²,

Hu³

2009

Electrochimica Acta

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Sugar-Sensitive Thin Films Composed of Concanavalin A and Glycogen

Cited by 27 publications

References 11 publications

Protein adsorption drastically reduces surface‐enhanced Raman signal of dye molecules

Protein adsorption drastically reduces surface‐enhanced Raman signal of dye molecules

Sugar-Sensitive Thin Films Composed of Concanavalin A and Sugar-Bearing Polymers

Loading of myoglobin into layer-by-layer films assembled by concanavalin A and dextran based on their biospecific recognition: An electrochemical study

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