Christof M Niemeyer

..Eventually, IPCR crossed the border from being a research method to a routine laboratory technique, enabling a broad range of applications in immunological research and clinical diagnostics...

..Hence, DNA can be used as a scaffold for positioning fluorescent proteins, as well as traditional fluorophores, with nanometer accuracy and shows great potential for use in the future of nanobiotechnology...

..The results are protein microarrays that can be used for the multiplexed detection of various antigens...

..Moreover, as the spatial selectivity of target immobilization is determined by the specificity of DNA base pairing, the assay is particularly suited for miniaturized microfluidics and lab-on-a-chip devices...

..Examples include the fabrication of functional biometallic nanostructures from gold nanoparticles and antibodies, applicable as diagnostic tools in bioanalytics...

..Moreover, we demonstrate that a pair of nanocircles, containing complementary oligonucleotide moieties, can be hybridized to form specific dimers, thereby generating a novel type of supramolecular DNA-protein nanostructures...

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..To this end, recent advances in cytochrome P450 development are reported and commonly used screening methods are surveyed...

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..The empirical results are capitalized on for the assembly of a CdS-TEV nanohybrid, which shows significantly higher performance as a photocatalytic mediator for fatty acid hydroxylation by CYP152A1 than bare CdS nanoparticles...

..Taking into account the greater versatility and convenience of handling of the self-assembly approach, this study demonstrates that DDI is an advantageous alternative for generating versatile and robust protein arrays...

..Azide-functionalized Ras protein was immobilized on a phosphane-modified surface by means of Staudinger ligation providing a highly chemoselective ligation method for the immobilization of proteins...

..Use of a confocal laser microscope with a UV laser as UV-light source enabled further reduction of biotin feature size opening access to nanostructured biochips...

..The psDNA-STV therefore hold particular promise for applications e.g. in proteome research and novel biosensing devices, where interfering endogenous nucleic acids need to be removed from analytes by nuclease digestion...

..Since this study demonstrates that biological fluorophores can be selectively incorporated into and optically coupled with nanoparticle-based devices, applications in DNA-based nanofabrication can be foreseen...

..In both cases, a comparable ELISA was 1000-fold less sensitive...

..As the consequence of the assay-based optimization, the ligation of MBP with the oligonucleotide was improved to near quantitative yields...

..Therefore, the self-assembly and regeneration capabilities of this approach should open the door to the generation of arrays of redox-enzyme devices for the screening of enzymes and their effectors...

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..These results suggest that IPCR has the potential to become a standard methodology in immunogenicity testing...

..The kinetic measurements revealed a significant increase in the reactivity of the complexes, in which GOX and HRP were immobilized in direct proximity on a complementary DNA carrier...

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..The hybridization capabilities are quantified in a microplate assay...

..This Review gives an overview on historical developments and the current state-of-the-art on apoenzyme reconstitution...

..We observed a detection limit of 320pg/ml of HBsAg using the M-IPCR, which was about 100-fold more sensitive than the analogous Magneto-ELISA, established in parallel for comparison purposes...

..Owing to slow temperature ramping, the entire assay requires about 17 h. The protocol allows to simultaneously analyze up to 384 samples with only 30 microl sample volume each...

..In general, our detailed description of fatty acid- and organic solvent-effects is of practical interest because it illustrates that optimization of modulators and cosolvents can lead to significantly increased yields in biocatalysis...

..Therefore, the microplate FRET method readily provides insights into the assembly process of complex DNA superstructures to verify and complement theoretical design approaches...

..The collected data provided the basis for the valid routine quantification of patient samples for the calculation of the pharmacokinetic behaviour of Aviscumine in patient plasma...

..Therefore, this analytical approach is useful to elucidate bioconjugate species, formed in the course of non-covalent synthesis of nanoparticle-biomolecule hybrid systems...

..The results demonstrate that this addressable chip is well suited for the generation of fluidically coupled multi-enzyme microreactors...

..Enzyme activity measurements not only indicated undisturbed biofunctionality of the tethered DNA but also enabled magnetic separation-based enrichment and recycling of the photoactivatable biocatalyst...

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..We demonstrate the performance of our system by parallel detection of cytokines in a multiplex immunoassay, employing silver deposition labeling and optical read-out...

..This Minireview highlights selected developments in the application of protein biochips in these fields...

..Therefore, qIPCR offers novel opportunities for the biomedical analysis of, for instance, neurodegenerative diseases and viral infections as well as new tools for the development of novel pharmaceuticals...

..These studies are paving the way for future applications in the self-assembly of photoactive supramolecular complexes, such as artificial light-harvesting systems...

..Hybridization efficiency of DNA-STV-conjugates depends on the position of the hybridization site in the carrier sequence, where the positions nearest to and farthest from the microarray surface proved to be most favorable...

..We anticipate that our concept will enable many applications, such as the generation of large arrays of surface-bound, recombinant proteins assembled by means of DNA-directed immobilization...

..This protocol significantly reduces handling time and costs of analysis...

..We anticipate that further refinement of this approach will enable applications, for instance, the assembly of ordered layers useful as transducers in biosensing...

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..This Review is intended to provide an overview of this fascinating and rapidly growing field of research from the structural design point of view...

..This critical review provides an overview on chemical approaches to the synthesis of DNA-protein conjugates and their applications in biomolecular nanosciences (96 references)...

..Such hybrid conjugates are also used for the biofunctionalization of planar surfaces for micro- and nanoarrays, and for decorating inorganic nanoparticles to enable applications in sensing, materials science, and catalysis...

..Strategies for creating protein patterns (as opposed to protein arrays) are also outlined. An outlook on promising and challenging future directions for protein biochip research and applications is also offered...

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..The values obtained for k(cat) (78.2+/-20.6 min(-1)) and K(M) (9.2+/-4.3 mM) indicated an approximately 100-fold increased catalytic activity over previously reported results...

..Subsequently, the conjugates are applied as molecular linkers in the DNA-directed immobilization of a biotinylated enzyme on a microplate, containing complementary capture oligonucleotides...

..The linear sensitivity range of the assay covered more than five orders of magnitude. Repeated measurements of rViscumin resulted in a mean standard deviation value of 14.2%...

..Overall, the results of this study suggested that the P450(BSbeta) enzyme represents a powerful catalyst in terms of the catalytic activity and operational stability...

..We anticipate many applications of our approach, ranging from protein microarrays to the arising field of nanobiotechnology...

..This behaviour coincides with the occurrence of an immobile band previously observed in electrophoretic experiments, indicating the formation of extremely large DNA-STV aggregate networks...

..The examples include the detection of viral and bacterial antigens, tumor markers, toxins, pathogens, cytokines and other targets in different biological sample materials...

..The performance of dendrimer-based DNA microarrays in the discrimination of SNPs is demonstrated...

..Rotavirus positive and negative stool samples were correctly identified by IPCR with a clear separation even of a 10,000-fold dilution of the positive stool samples from the negative control...