Optimization of immobilization

Raicevic, S., Plecas, I., Lalovic, D. I. Velikovic, V. 1999. Optimization of immobilization of strontium and uranium by the solid matrix. Materials Research Society Symposium Proceedings, 556, 135-142. 

The search for new approaches which can give increasing sensitivity to S PR-based biosensors is associated with the optimization of immobilization of the biological components on the transducer surface. 

Immobilization. Enzymes, as individual water-soluble molecules, are generally efficient catalysts. In biological systems they are predorninandy intracellular or associated with cell membranes, ie, in a type of immobilized state. This enables them to perform their activity in a specific environment, be stored and protected in stable form, take part in multi-enzyme reactions, acquire cofactors, etc. Unfortunately, this optimization of enzyme use and performance in nature may not be directiy transferable to the laboratory. 

When selecting a suitable feed symp, the main criteria are optimization of enzyme productivity and minimization of the formation of by-products. Typical feed symp specifications are shown in Table 5. Higher symp concentration and higher viscosity results in a reduced isomerization rate due to diffusion resistance in the pores of the immobilized enzyme. A deaeration step is desirable to remove dissolved oxygen that would otherwise iacrease the formation of by-products. The pH is adjusted to the optimum level for the productivity of the enzyme. 

An intriguing influence of a cosolvent immiscible with water on the enantioselec-tivity of the enzyme-catalyzed hydrolysis was observed. It was proven that enzyme enantioselectivity is directly correlated with the cosolvent hydrophobicity. In the best example, for ethyl ether as cosolvent, the reaction proceeded with E = 55, and the target compound was obtained in 33% yield with 92.7% ee. This finding may be of great practical importance, particularly in industrial processes [24], since it will enable better optimization of enzyme-catalyzed processes. It is clear that, in future, immobilized enzymes, as heterogeneous catalysts, wiU be widely used in most industrial transformations, especially in the preparation of pharmaceuticals [25]. 

The choice of immobilization strategy obviously depends on the enzyme, electrode surface, and fuel properties, and on whether a mediator is required, and a wide range of strategies have been employed. Some general examples are represented in Fig. 17.4. Key goals are to stabilize the enzyme under fuel cell operating conditions and to optimize both electron transfer and the efficiency of fuel/oxidant mass transport. Here, we highlight a few approaches that have been particularly useful in electrocatalysis directed towards fuel cell applications. 

Recent reports describe the use of various porous carbon materials for protein adsorption. For example, Hyeon and coworkers summarized the recent development of porous carbon materials in their review [163], where the successful use of mesoporous carbons as adsorbents for bulky pollutants, as electrodes for supercapacitors and fuel cells, and as hosts for protein immobilization are described. Gogotsi and coworkers synthesized novel mesoporous carbon materials using ternary MAX-phase carbides that can be optimized for efficient adsorption of large inflammatory proteins [164]. The synthesized carbons possess tunable pore size with a large volume of slit-shaped mesopores. They demonstrated that not only micropores (0.4—2 nm) but also mesopores (2-50 nm) can be tuned in a controlled way by extraction of metals from carbides, providing a mechanism for the optimization of adsorption systems for selective adsorption of a large variety of biomolecules. Furthermore, Vinu and coworkers have successfully developed the synthesis of... 

A flow injection optical fibre biosensor for choline was also developed55. Choline oxidase (ChOX) was immobilized by physical entrapment in a photo-cross-linkable poly(vinyl alcohol) polymer (PVA-SbQ) after adsorption on weak anion-exchanger beads (DEAE-Sepharose). In this way, the sensing layer was directly created at the surface of the working glassy carbon electrode. The optimization of the reaction conditions and of the physicochemical parameters influencing the FIA biosensor response allows the measurement of choline concentration with a detection limit of 10 pmol. The DEAE-based system also exhibited a good operational stability since 160 repeated measurements of 3 nmol of choline could be performed with a variation coefficient of 4.5%. 

Peluso P., Wilson D.S., Do D., Tran H., Venkatasubbaiah M., Quincy D., Heidecker B., Poindexter K., Tolani N., Phelan M., Witte K., Jung L.S., Wagner P., Nock S., Optimizing antibody immobilization strategies for the construction of protein microarrays, Anal Biochem. 2003 312 113-124. 

Our results have shown that the use of STARBURST dendrimers for the covalent coupling of the molecules of biological interest can lead to a reproduc-ibly performing product with very little impact on the biological activity of the immobilized protein. By careful optimization of the reaction parameters, it has... 

Model system for testing utilization of immobilized affinity peptides - SMC (CIM ) optimally present small affinity ligands Ligand utilization, comparison with agarose, cellulose and synthetic particle based polymers Affinity (peptides) disks [85]... 

Since the large-scale application of immobilized enzymes in the 1960s, substantial research efforts have aimed to optimize the structure of carrier materials for better catalytic efficiency. To date, nanoscale materials may provide the upper limits in... 

After the optimization of these conditions, by adding an azide to the input stream it was possible to synthesize a range of substituted triazoles in a heterogeneously catalysed three-component reaction (Scheme 18). After the CFC, the stream was passed through a column containing a resin-immobilized copper-based catalyst, which was used in a previous work by the same authors to successfully catalyze the formation of triazoles from alkynes and azides [44]. An immobilized thiourea-containing cartridge was subsequently used to remove any leached Cu catalyst. In a similar way as for the alkynes production, the series of resins was used to purify the product. 

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