Immobilization proteins

Covalent immobilization of proteins on nricrostmctured gold surfaces was studied in [226]. On Arese substrates, which were prepared by pCP aird etching. Are immobilization sites of proteins could be spatially controlled using air amino-reactive SAM. The whole process, i.e. production of Are micropattemed substrate including SAM exchairge aird protein immobilization, took a reasonably small amount of time ( 24 h), providing some flexibility in the experimental work. 

Ramsden J J 1998 Biomimetic protein immobilization using lipid bilayers Biosensors Bloelectronics 13 593—8... 

Traditional appHcations for latices are adhesives, binders for fibers and particulate matter, protective and decorative coatings (qv), dipped goods, foam, paper coatings, backings for carpet and upholstery, modifiers for bitumens and concrete, and thread and textile modifiers. More recent appHcations include biomedical appHcations as protein immobilizers, visual detectors in immunoassays (qv), as release agents, in electronic appHcations as photoresists for circuit boards, in batteries (qv), conductive paint, copy machines, and as key components in molecular electronic devices. 

Two-dimensional protein layer orientation could be also effected by metal-ion coordination Monolayer of iminodiacetate-Cu(II) lipid was successfully employed as substrate for oriented immobilization of proteins naturally displaying histidine residues on their surface [37]. Affmity-resin-displaying Ni(II) complexes could also be successfully employed for oriented protein immobilization [38]. 

Monolithic silica columns with various surface derivatization, including ion exchange (Xie et al., 2005), and chiral functionalities (Chen et al., 2002 Lubda et al., 2003 Chankvetadze et al., 2004), as well as protein-immobilized monoliths (Kato et al., 2005) have been reported. The latter will be an important part of an integrated multidimensional separation/identification system. 

Houston B, Peddie D (1989) A method for detecting proteins immobilized on nitrocellulose membranes by in situ derivatization with fluorescein isothiocyanate. Anal Biochem 177 263-267... 

In this chapter, immobilization of proteins on these mesoporous silicas and PMO is first introduced, followed by a description of protein immobilization on mesoporous carbon materials. The adsorption behavior of other biopolymers such as... 

In addition to popular mesoporous silica materials, mesoporous silica supports with various morphologies have also been used for protein immobilization. Tang and coworkers synthesized lotus-leaf-like silica flakes with a three-dimensionally connected nanoporous structure and controllable thickness, which were used for immobilization of ribonuclease A [126]. The synthesized silica flakes have a thickness of200 nm, and a diameter of 3 mm, showing a much higher initial adsorbing rate of... 

For application of protein-immobilized porous materials to sensor fields, use of an electroactive substance as the framework material is important. DeLouise and Miller demonstrated the immobilization of glutathione-S-transferase in electrochemically etched porous silicon films [134], which are attractive materials for the construction of biosensors and may also have utility for the production of immobilized enzyme bioreactors. Not limited to this case, practical applications of nanohybrids from biomolecules and mesoporous materials have been paid much attention. Examples of the application of such hybrids are summarized in a later section of this chapter. 

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... 

Kumar, C.V. and Choudhari A. (2000) Proteins immobilized at the galleries of layered alpha-zirconium phosphate structure and activity studies. Journal of the American Chemical Society, 122, 830-837. 

Modified dextrans, such as carboxymethylated dextran were also coated onto glass slides to serve as matrix for protein immobilization and at the same time reduce non-specific binding22. 

Kumada Y, Shiritani Y, Hamasaki K, Ohse T, Kishimoto M (2009) High biological activity of a recombinant protein immobilized onto polystyrene. Biotechnol J 4 1178-1189... 

Hahn, C.D., Leitner, C., Weinbrenner, T., Schlapak, R., Tinazli, A., Tampe, R., Lackner, B., Steindl, C., Hinterdorfer, P., Gruber, H.J., and Holzl, M. (2007) Self-assembled monolayers with latent aldehydes for protein immobilization. Bioconjugate Chem. 18, 247-253. 

Stolowitz, M.L., Ahlem, C., Hughes, K.A., Kaiser, R.J., Kesicki, E.A., Li, G., Lund, K.P., Torkelson, S.M., and Wiley, J.P. (2001) Phenylboronic acid-salicylhydroxamic acid bioconjugates. 1. A novel boronic acid complex for protein immobilization. Bioconjugate Chem. 12, 229-239. 

Sun, X.-L., Stabler, C.L., Cazalis, C.S., and Chaikof, E.L. (2006) Carbohydrate and protein immobilization onto solid surfaces by sequential diels-alder and azide-alkyne cycloadditions. Bioconjugate Chem. 17, 52-57. 

M. Vladimir, M. Mirsky, O. Riepl, and O. Wolfbeis, Capacitive monitoring of protein immobilization and antigen—antibody reactions on monomolecular alkylthiol films on gold electrodes. Biosens. Bioelectron. 12, 977-989 (1997). 

At present, a wide range of solid substrates are available for protein immobilization. According to the protein attachment strategies, namely, adsorption, affinity binding, and covalent binding, all these substrates can be separated into three main parts. Surfaces like ploy(vinylidene fluoride) (PVDF), poly(dimethylsiloxane) (PDMS), nitrocellulose, polystyrene, and poly-1-lysine coated glass can adsorb proteins by electrostatic or hydrophobic forces. A potential drawback of such substrates is the difficulty... 

W.H. Scouten, J.H. Luong, and R.S. Brown, Enzyme or protein immobilization techniques for applications in biosensor design. Trends Biotechnol. 13,178—185 (1995). 

The physical adsorption of protein onto the surface of an electrode is a simple immobilization method. The adsorption is obtained by volatilizing the buffers containing proteins. The physical adsorption needs no chemical reagent, seldom activation and rinse, so that the bioactivities of the immobilized proteins can be retained well. However, the immobilized proteins are easy to break off from the electrode, which restrict broad applications of this method. Below are some examples of the physical adsorption of proteins immobilized on electrodes. 

Suaud-Chagny and Gonon [3] presented a new procedure for protein immobilization adapted to carbon microelectrode characteristics. The principle of this method of immobilization is based on the association of the protein with an inert porous film immobilized around the active tip of the electrode. For this purpose the carbon was coated with an inert, electrochemically obtained protein sheath (bovine serum albumin, BSA) a few micrometers thick. Then the sheath around the fiber was impregnated with lactate dehydrogenase (LDH), which could be immobilized onto the electrode and resulted in an electrode sensitive to pyruvate. 

J.J. Feng, G. Zhao, J.J. Xu, and H.Y. Chen, Direct electrochemistry and electrocatalysis of heme proteins immobilized on gold nanoparticles stabilized by chitosan. Anal. Biochem. 342, 280-286 (2005). 

Chen RJ, Zhang Y G, Wang DW, Dai HJ(2001b)N oncovalent sidewall functionalization of single-walled carbon nanotubes for protein immobilization. J. Am. Chem. Soc. 123 3838-3839. 

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