Carbodiimide, surface attachment

For the determination of these compounds a binding inhibition immunoassay, consisting of the competitive immunoreaction of the unbound antibody present in an analyte-antibody mixture with the hapten derivative immobilized at the sensor surface, has been applied. With the aim of assuring the regeneration and reusability of the surface without denaturation of the immobilized molecule, the formation of an alkanethiol monolayer was carried out to provide covalent attachment of the ligand to the functionalized carbodiimide surface in a highly controlled way. For DDT, the assay sensitivity was evaluated in the 0.004 - 3545 pg/l range of pesticide concentration by the determination of the limit of detection 0.3 pg/1 and the I50 value 4.2 pg/1. 

Carbodiimides are widely used to mediate the attachment of biomarkers to polypeptides. Examples include carbodiimides with ferrocenyl substituents. Also, peptides are covalently modified with ferrocenecarboxylic acid using EDCCl and N-hydroxy-succinimide to promote the coupling to surface lysines. They also mediate the attachment of substituents to single walled nanotubes (SWNTs) and multiwalled nanotubes (MWNTs). Also, microdots are attached to virus molecules using a water soluble carbodiimide. The attachment of viral DNA to gold particles is used in the manufacture of a new type of vaccine. 

Andres and coworkers demonstrated the iCP of densely packed alkanethiolate-functionalized Au nanoparticle arrays in monolayer and multilayer structures.81,82 Dense and hexagonally packed monolayers of nanoparticles were first assembled on a water surface. By using the Langmuir-Schafer technique, the Au nanoparticle monolayer was transferred to a PDMS stamp, and printed onto a substrate. Multilayers were prepared by repeating the printing process in an LbL scheme, in which subsequent particle layers may be made up of the same or different types of particles. Similarly, the assembly of irregular, densely packed monolayers of polystyrene nanoparticles on iCP substrates via carbodiimide coupling was reported.83 The conformal contact of the carbodiimide-functionalized polystyrene particles resulted in the covalent attachment of the nanoparticles at a carboxylate-functionalized surface. 

Several useful schemes for attaching nucleic acid probes onto electrode surfaces have thus been developed [2-8]. The exact immobilization protocol often depends on the electrode material used for signal transduction. Common probe immobilization schemes include attachment of biotin-functionalized probes to avidin-coated surfaces [15], self-assembly of organized monolayers of thiol-functionalized probes onto gold transducers [16], carbodiimide covalent binding to an activated surface [17], as well as adsorptive accumulation onto carbon-paste or thick-film carbon electrodes [15-30]. 

Covalent, random Epoxy, aldehyde, carbodiimide groups React with primary amines of lysine and arginine High densihes are available and strong protein attachment/ random orientahon and surface interference... 

Organothiols chemisorbed on gold surfaces are especially usefiil for electrochemical and chemical studies. For example, carboxylic acid terminated monolayers are often used. An example is the attachment of cytochrome C with its side chain lysine groups to acid terminated self assembled monolayer gold electrodes. Carbodiimide mediated esterification of ferrocene-l,l -dicarboxylic acid with phenols is used to construct thermotropic liquid crystals. ... 

The most employed method for developing stable materials for biosensor application is via covalent attachment. There are two routes to carry out this process (i) the first involves direct immobilization of biomolecules onto the transducer surface that contains residual groups (carboxyl, aldehyde, hydroxyl). Despite its simplicity, this procedure is characterized by poor reproducibility, (ii) The second and mostly used route involves the derivatiza-tion of the transducer smface with a cross-linker such as glutaraldehyde, carbodiimide/succinimide or avidin/biotin pathways [27]. Commercial preactivated membranes are available and can be utilized as support to immobilize biomolecules. 

Various approaches were probed to attach the molecule of cytochrome c to the surface of the monolayers. Covalent attachment could be achieved by carbodiimide coupling of the acid groups on the monolayer surface with the amine groups of the protein356,380. Electrostatic adsorption of the protein on the surface of acid-terminated monolayers381,382 was achieved by Tarlov and co workers. Cytochrome c also adsorbs on the surface of a pyridine-containing monolayer383. 

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