کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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1165568 | 1491106 | 2012 | 8 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Improvement of the homogeneity of protein-imprinted polymer films by orientated immobilization of the template Improvement of the homogeneity of protein-imprinted polymer films by orientated immobilization of the template](/preview/png/1165568.png)
A method for preparing homogeneous protein-imprinted polymer films with orientated immobilization of template is described. The template methyl parathion hydrolase (MPH) was modified with a peptide linker (Gly-Ser)5–Cys and was immobilized on a cover glass with a fixed orientation via the linker. The activity of the fusion enzyme (MPH-L) was evaluated by determining the product's absorbance at 405 nm (A405). Both the free and the immobilized MPH-L showed higher retention of the bioactivity than the wide type enzyme (MPH-W) as revealed by the A405 values for MPH-Lfree/MPH-Wfree (1.159/1.111) and for MPH-Limmobilized/MPH-Wimmobilized (0.348/0.118). The immobilized MPH-L also formed a more homogeneous template stamp compared to the immobilized MPH-W. The molecularly imprinted polymer films prepared with the immobilized MPH-L exhibited high homogeneity with low Std. Deviations of 80 and 200 from the CL intensity mean volumes which were observed for batch-prepared films and an individual film, respectively. MPH-L-imprinted polymer film also had a larger template binding capacity indicated by higher CL intensity mean volume of 3900 INT over 2500 INT for MPH-W-imprinted films. The imprinted film prepared with the orientated immobilization of template showed an imprinting factor of 1.7, while the controls did not show an imprinting effect.
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► MPH was genetically modified at its C-terminal with (Gly-Ser)5–Cys.
► MPH-L was immobilized with fixed orientation via disulfide chemistry.
► The immobilized MPH-L retained the activity of MPH.
► MPH-L formed a homogeneous template.
► Homogeneous MIP film was obtained with orientated immobilization of the template.
Journal: Analytica Chimica Acta - Volume 726, 13 May 2012, Pages 85–92