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The characteristics of the azopolymer make it possible to immobilize a wide variety of biological molecules on the same substrate through a one-step photoirradiation process To take advantage of the functionality of these biomolecules, identifying an immobilization process that does not lead to deactivation of the molecules is important In particular, biomolecules such as proteins show sensitive behavior in terms of changes in environment, as shown by the denaturing of proteins when the surrounding temperature increases even slightly Since it is possible that damage to biomolecules following photoinduced immobilization could trigger functional degradation, the authors rst examined the activity of an immobilized enzyme An aqueous solution of 1-mg/mL bacterial protease (subtilisin; 275 kDa, Sigma) was spotted onto the surface of an azopolymer, and the surface was irradiated with a laser beam of 488-nm wavelength and 80-mW/cm2 optical power density for 5 min to immobilize the enzyme.

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As a control experiment, a similar specimen was prepared without photoirradiation The activity of the subtilisin was veri ed as the hydrolysis of the arti cial substrate (tert-butoxycarbonyl-Gly-Gly-Leu-p-nitroanilide, Mw=4655, Merck) The arti cial substrate solution was spotted onto the azopolymer surface in the same area where the subtilisin had been immobilized, and then the specimen was maintained at 37 1C and 85% relative humidity for 1 h The hydrolysis of the arti cial substrate was determined spectroscopically by immediately measuring the absorbance of the reactant at a wavelength of 410 nm The conversion ratio of the reaction was B10% for the subtilisin-immobilized sample, whereas it was B1% for the control sample (without photoirradiation) These results clearly show that biomolecules immobilized on an azopolymer surface can maintain their enzyme functionality during and after the immobilization process.

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Are there KIP s for social media Some are starting to emerge. Take for example, the Econsultancy blog post by Chris Lake at http://econsultancy.com/blog/4887-35-socialmedia-kpis-to-help-measure-engagement called 35 social media KPIs to help measure engagement. Chris offers some clear insights about social media measurement and then provides the following:

Next, the authors investigated how deformation of an azopolymer surface can be induced by biomolecules as well as by microspheres A phosphate-buffered saline (PBS) solution containing Cy-5-linked immunoglobulin, IgG, was spotted onto the surface of an azopolymer After evaporating the solution, the surface was irradiated for 30 min with light of 470-nm wavelength and 10-mW/cm2 optical power density from an array of blue light-emitting diodes (LEDs) and then the.

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surface was washed for 30 min with PBS containing 0.01-wt% Tween 20 as a nonionic surfactant. The amount of immobilized IgG was con rmed by the uorescence intensity of the spot, and the minimum detectable amount was 10 pg. Next, a surface image was obtained by tapping mode AFM (Digital Instruments, Dimension 3100) using a sharp silicon cantilever with a tip radius of o5 nm. In Fig. 9.9a, the azopolymer surface is covered with a layer of small granulated particles of 10 30 nm diameter and B8 nm height, where the height was estimated from the defects and the edge of the layer. The sizes of the particles were nearly equivalent to one subunit of IgG (B10 nm), considering that the image includes AFM tip convolution artifacts. The layer is so at that the IgG monolayer is believed to be located on the azopolymer surface. The sample was subsequently washed with PBS containing 2-wt% sodium dodecyl sulfate to remove the IgG. After con rming that the uorescence from the spot had disappeared, another AFM image was obtained, which is shown in Fig. 9.9b. Dents of B20 nm diameter

Azopolymer (a) Size of antibody (15 nm)

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Azopolymer (b)

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