Quantitative analysis of protein adsorption on a planar surface by Fourier transform infrared spectroscopy: lysozyme adsorbed on hydrophobic silicon-containing polymer

Yokoyama, Yasuhiro; Ishiguro, R.; Maeda, Hirotaka; Mukaiyama, Mayumi; Kameyama, Keiichi; Hiramatsu, Kazumasa

doi:10.1016/j.jcis.2003.07.011

Cited by 30 publications

(35 citation statements)

References 44 publications

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“…Comparing the number of peptides bound to the IMAC chips to a monolayer of peptides, that is, 0.1-0.5 mg cm À2 , [13,14,31] we see that it by far exceeds one monolayer and, as expected, corresponds to a multilayer situation.…”

Section: à2supporting

confidence: 67%

“…[4,7,12,14,30] However, this leads to a smaller number of bound peptides and thereby a lower ATR signal (or any measurement signal). In ATR spectroscopy, this can be compensated for by a higher number of internal reflections of the IRE, which always increase the signal-to-noise ratios, [5] but increases the complexity of the set-up and, for instance, limits the selection and possible combinations of IRE material.…”

Section: à2mentioning

confidence: 99%

“…conformational change upon adsorption or molecular interaction, [14][15][16] protein secondary structure determination, [7,[17][18][19] and orientation in proteins and lipids. [7,20,21] Very recently some progress has also been made to functionalise IREs with crystals based on germanium [3,4,22] and silicon.…”

Section: Introductionmentioning

confidence: 99%

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A Novel ATR‐FTIR Approach for Characterisation and Identification of Ex Situ Immobilised Species

et al. 2007

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We demonstrate a novel method to analyse ex situ prepared protein chips by attenuated total reflection Fourier IR spectroscopy (ATR‐FTIR), which circumvents tedious functionalisation steps of internal reflection elements (IREs), and simultaneously allows for complementary measurements by other analytical techniques. This concept is proven by utilising immobilised metal affinity capture (IMAC™) chips containing about 10 μm thick films of copolymers coated with nitrilotriacetic acid (NTA) groups, which originally was manufactured for surface enhanced laser desorption ionisation (SELDI) spectrometry. Three immobilisation steps were analysed by ATR‐FTIR spectroscopy: 1) NTA complexation with nickel(II) ions 2) binding of two histidine (His)‐tagged synthetic peptides of 25 (25‐His6) and 48 (48‐His6) amino acids to the NTA‐groups and 3) attachment of a ligand, mesyl amide, to the surface‐bound 48‐His6. Despite interference from H2O, both amide I and II were well resolved. Utilising peptide adsorption in the thick copolymer matrix yields a high saturation peptide concentration of ≈100 mg mL−1 and a dissociation constant of 116±11 μM, as determined by a detailed analysis of the Langmuir adsorption isotherm. The mesyl amide ligand was directly seen in the raw ATR‐FTIR spectrum with specific peaks in the fingerprint region at 1172 and 1350 cm−1. Several aspects of the fine structure of the amide I band of the peptide were analysed: influences from secondary structure, amino side chains and competing contamination product. We believe that this approach has great potential as a stand‐alone or complementary analytical tool for determination of the chemical composition of functionalised surfaces. We emphasise further that with this approach no chemical treatment of IREs is needed; the chips can be regenerated and reused, and applied in other experimental set‐ups.

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Section: à2supporting

confidence: 67%

Section: à2mentioning

confidence: 99%

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A Novel ATR‐FTIR Approach for Characterisation and Identification of Ex Situ Immobilised Species

et al. 2007

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“…More recently, Yokoyama et al (2003) used this technique to study the adsorption of hen egg white lysozyme onto a solid polytris(trimethylsiloxy) silylstyrene surface, while Xie et. al.…”

Section: Introductionmentioning

confidence: 99%

Adsorption of human immunoglobulin G to poly ( β-hydroxybutyrate) (PHB), poly (L- lactic Acid) (PLLA) and PHB/PLLA blends

Lima

Duek

Santana

2009

Braz. J. Chem. Eng.

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-Biodegradable polymeric materials can be used as temporary implants and may be able to carry out specific functions for a pre-determined period prior to their degradation in vivo. In order to be used they must present characteristics of biocompatibility. When a material comes into contact with blood, the instantaneous adsorption of protein occurs on its surface. Coverage of the surface by γ-globulins causes the adhesion of platelets and, consequently, provokes the process of blood coagulation. In this context, the objective of the present paper was to quantify the adsorption of human Immunoglobulin G (HIgG) to poly (β-hydroxybutyrate) (PHB), poly (L-lactic acid) (PLLA) and PHB/PLLA blends using the FTIR/ATR technique. The results showed the occurrence of HlgG adsorption for all three systems, PHB having the highest adsorption density, about Γ max =2.57μg/cm 2 , followed by PHB/PLLA blends (Γ max =0.85μg/cm 2 ) and PLLA with the lowest values (Γ max =0.25μg/cm 2 ).

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“…In our previous study, we used Fourier transform infrared (FTIR) spectroscopy with attenuated total reflection (ATR) to study the adsorption of hen egg white lysozyme dissolved in D 2 O onto the surface of solid poly tris(trimethylsiloxy)-silylstyrene (pTSS) to measure infrared spectra of a pTSS film prepared on an ATR prism in D 2 O solutions containing lysozyme [21]. We derived accurate spectra attributed to only the protein adsorbed on the pTSS surface by subtracting the contributions of pTSS, solvent, and unbound lysozyme from the whole spectra.…”

Section: Introductionmentioning

confidence: 99%

Modes of conformational changes of proteins adsorbed on a planar hydrophobic polymer surface reflecting their adsorption behaviors

Ishiguro

Yokoyama

Maeda

et al. 2005

Journal of Colloid and Interface Science

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Quantitative analysis of protein adsorption on a planar surface by Fourier transform infrared spectroscopy: lysozyme adsorbed on hydrophobic silicon-containing polymer

Cited by 30 publications

References 44 publications

A Novel ATR‐FTIR Approach for Characterisation and Identification of Ex Situ Immobilised Species

A Novel ATR‐FTIR Approach for Characterisation and Identification of Ex Situ Immobilised Species

Adsorption of human immunoglobulin G to poly ( β-hydroxybutyrate) (PHB), poly (L- lactic Acid) (PLLA) and PHB/PLLA blends

Modes of conformational changes of proteins adsorbed on a planar hydrophobic polymer surface reflecting their adsorption behaviors

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