Physical entrapment methods

When immobilizing biocatalysts within polymer gels using physical entrapment methods, we may take advantage of the great resistance to the diffusion of macromolecular substances due to the gel porosity. However, this limited diffusion within the gel phase also causes a reduced mass transfer rate for low... 

However, Yokoyama et al. (1994) reported that a ratio between the chemically conjugated and physically entrapped ADR (i.e., DOX) was not determined, and considerable amounts of adriamycin derivatives formed and were incorporated in the micelles. Yokoyama et al. (1998) quantitatively measured the ADR incorporated in the inner core using the improved synthetic method, and analyzed the effects of the ADR contents (both by chemical conjugation and physical entrapment) on micelle stability andn vivo antitumor activity. The copolymer used by Yokoyama et al. (1998) was polyethylene glycol)-fepoly(aspartic acid) block copolymer (PB P(Asp)). The chemical conjugation and physical entrapment methods can be referred to in the earlier section of preparation of polymeric micelles. 

Table 1. Various factors affecting the degree of immobilization in the physical entrapping method...

Incorporation of uricase into CPs nanotubes for measuring the urea concentration has been achieved [169]. Urease was immobilized by a physical entrapment method. The sensor displayed a linear concentration range of urea between 1.22 pM and 3.85 mM. The sensitivity and detection limits... 

The first two methods have the advantage that no modification of the homogeneous catalyst is needed. Surface hydrogen-bonded catalysts are limited to cationic complexes, while physical entrapment is more widely applicable. However, both methods are very sensitive to the solvent properties of the reaction medium. The chemical methods of immobilization require modification of the ligand, and this may be quite laborious. In the case of irreversible catalyst deacti-... 

Synthetic methods include the use of silanes bearing a chiral group for silylating the surface of the porous sol-gel silica, the use of such silanes as monomers or co-monomers in the polycondensation, the physical entrapment of chiral molecules, the imprinting of SG materials with chiral templates and the creation of chiral pores, and the induction of chirality in the matrix skeleton itself 48... 

Because enzymes present such an attractive possibility for achieving chemical selectivity, enzyme electrodes were the first enzymatic chemical sensors (or first biosensors) made. The early designs used any available method of immobilization of the enzyme at the surface of the electrode. Thus, physical entrapment using dialysis membranes, meshes, and various covalent immobilization schemes have been... 

In this polymerization, the biofunctional component (enzyme) can be concentrated in an interfacial area between the frozen ice crystal and the supercooled monomer phase, and immobilized by molecular entanglement between the enzyme and polymer molecules. This is a different procedure for fixation from the usual entrapping method with a crosslinked structure in a gel. Therefore, we may call this procedure the adhesion-method to distinguish it from the usual entrapping. This term was extended to cover the use of the usual synthetic polymers including hydrophobic polymers as the supports. One of the characteristic properties of products obtained in this way was that there is a maximum activity at a certain monomer concentration. The maximum activity is observed when the increased inner surface area is balanced by the increased leakage of enzyme and these occur with a decrease of monomer concentration. Immobilization by physical entrapping was also studied by Rosiak [26], Carenza [27] and Ha [28]. 

Entrapping I Encapsulation Methods Enzymes can be immobilized by physical entrapment without involving any chemical bonding. This route is somewhat more laborious but yields an enzyme that is least altered by the immobilization. 

Functionalization of surfaces with photopolymerized monomers or sol-gel matrices has also been used. In this case, the biomolecules can be physically entrapped within the supporting matrix. The method is applicable for almost all types of surfaces, it is easy to handle, and the large spectrum of monomer precursors commercially available permits the immobilization of basically all biological elements. In this case, the absence of chemical bond formation helps to preserve the activity of the bioreagent during the immobilization process. However, several drawbacks such as leaking of biocomponent and possible diffusion barriers restrict the performance of biosensor devices fabricated using this procedure. 

Physical immobilization methods do not involve covalent bond formation with the enzyme, so that the native composition of the enzyme remains unaltered. Physical immobilization methods are subclassified as adsorption, entrapment, and encapsulation methods. Adsorption of proteins to the surface of a carrier is, in principle, reversible, but careful selection of the carrier material and the immobilization conditions can render desorption negligible. Entrapment of enzymes in a cross-linked polymer is accomplished by carrying out the polymerization reaction in the presence of enzyme the enzyme becomes trapped in interstitial spaces in the polymer matrix. Encapsulation of enzymes results in regions of high enzyme concentration being separated from the bulk solvent system by a semipermeable membrane, through which substrate, but not enzyme, may diffuse. Physical immobilization methods are represented in Figure 4.1 (c-e). 

The behavior of immobilized enzymes differs from that of dissolved enzymes because of the effects of the support material, or matrix, as well as conformational changes in the enzyme that result from interactions with the support and covalent modification of amino acid residues. Properties observed to change significantly upon immobilization include specific activity, pH optimum, Km, selectivity, and stability.23 Physical immobilization methods, especially entrapment and encapsulation, yield less dramatic changes in an enzyme s catalytic behavior than chemical immobilization methods or adsorption. The reason is that entrapment and encapsulation result in the enzyme remaining essentially in its native conformation, in a hydrophilic environment, with no covalent modification. 

Both chemical and physical methods may be used to immobilize biocatalysts while retaining or modifying their activity, selectivity, or stability. Among the techniques used for immobilization of enzymes are physical adsorption, covalent bonding, ionic binding, chelation, cross-linking, physical entrapment, microencapsulation, and retention in permselective membrane reactors. The mode of immobilization employed for a particular application depends not only on the specific choice of enzyme and support, but also on the constraints imposed by the microenvironment associated with the application. 

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