DNA-Directed Immobilization: Efficient, Reversible, and Site-Selective Surface Binding of Proteins by Means of Covalent DNA–Streptavidin Conjugates

Niemeyer, Christof M.; Boldt, Larissa; Ceyhan, Bülent; Blohm, Dietmar

doi:10.1006/abio.1998.3017

Cited by 155 publications

(126 citation statements)

References 26 publications

Supporting

Mentioning

119

Contrasting

Unclassified

Order By: Relevance

“…In the last decade, nucleic acids have received increasing interest as bioreceptors for biosensor and biochip technologies [26][27][28][29][30][31]. The complementarity of adenine : thymine (A : T) and cytosine : guanosine (C : G) pairing in DNA (Fig.…”

Section: Nucleic Acidsmentioning

confidence: 99%

Biosensors and biochips: advances in biological and medical diagnostics

Vo‐Dinh

Cullum

2000

Fresenius' Journal of Analytical Chemistry

476

249

View full text Add to dashboard Cite

In the past two decades, the biological and medical fields have seen great advances in the development of biosensors and biochips capable of characterizing and quantifying biomolecules. This review is meant to provide an overview of the various types of biosensors and biochips that have been developed for biological and medical applications, along with significant advances over the last several years in these technologies. It also attempts to describe various classification schemes that can be used for categorizing the different biosensors and provide relevant examples of these classification schemes from recent literature.

show abstract

Section: Nucleic Acidsmentioning

confidence: 99%

Biosensors and biochips: advances in biological and medical diagnostics

Vo‐Dinh

Cullum

2000

Fresenius' Journal of Analytical Chemistry

476

249

View full text Add to dashboard Cite

show abstract

“…[18±36] We here report on the use of a software tool, DNASequenceGenerator, previously developed for DNA sequence design, [37] for generating a small library of DNA oligomers for the DNA-directed immobilization (DDI) [38,39] of proteins on a DNA microarray. This library, optimized in silico with respect to uniform hybridization efficiency and avoidance of nonspecific cross-hybridization, was experimentaly tested and compared in vitro using microarray-experiments with a library taken from literature.…”

Section: Introductionmentioning

confidence: 99%

Microarray‐Based in vitro Evaluation of DNA Oligomer Libraries Designed in silico

Feldkamp

Wacker

Schroeder³

et al. 2004

ChemPhysChem

Self Cite

View full text Add to dashboard Cite

We report on the microarray-based in vitro evaluation of two libraries of DNA oligonucleotide sequences, designed in silico for applications in supramolecular self-assembly, such as DNA computing and DNA-based nanosciences. In this first study which is devoted to the comparison of sequence motif properties theoretically predicted with their performance in real-life, the DNA-directed immobilization (DDI) of proteins was used as an example of DNA-based self-assembly. Since DDI technologies, DNA computing, and DNA nanoconstruction essentially depend on similar prereguisites, in particular, large and uniform hybridization efficiencies combined with low nonspecific cross-reactivity between individual sequences, we anticipate that the microarray approach demonstrated here will enable rapid evaluation of other DNA sequence libraries.

show abstract

“…In addition, the microarray can also be used for the addressable adsorption of other type of species with c-DNA tags, e.g., antibodies, 15,16 and membrane-residing proteins, which offers a stable, mild, and efficient immobilization procedure.…”

Section: Discussionmentioning

confidence: 99%

“…Consequently, DNA hybridization leads to self-sorting and immobilization of the vesicles to their respective array sites. This approach was inspired by the work of Niemeyer et al, 15,16 who used it to immobilize proteins to specific sites on a surface. The method offers stable tethering of the lipid vesicles to the surface while maintaining physiological conditions throughout the assay.…”

Section: Introductionmentioning

confidence: 99%

Addressable adsorption of lipid vesicles and subsequent protein interaction studies

et al. 2008

View full text Add to dashboard Cite

We demonstrate a convenient chip platform for the addressable immobilization of protein-loaded vesicles on a microarray for parallelized, high-throughput analysis of lipid-protein systems. Self-sorting of the vesicles on the microarray was achieved through DNA bar coding of the vesicles and their hybridization to complementary strands, which are preimmobilized in defined array positions on the chip. Imaging surface plasmon resonance in ellipsometric mode was used to monitor vesicle immobilization, protein tethering, protein-protein interactions, and chip regeneration. The immobilization strategy proved highly specific and stable and presents a mild method for the anchoring of vesicles to predefined areas of a surface, while unspecific adsorption to both noncomplementary regions and background areas is nonexistent or, alternatively, undetectable. Furthermore, histidine-tagged receptors have been stably and functionally immobilized via bis-nitrilotriacetic acid chelators already present in the vesicle membranes. It was discovered though that online loading of proteins to immobilized vesicles leads to cross contamination of previously loaded vesicles and that it was necessary to load the vesicles offline in order to obtain pure protein populations on the vesicles. We have used this cross-binding effect to our benefit by coimmobilizing two receptor subunits in different ratios on the vesicle surface and successfully demonstrated ternary complex formation with their ligand. This approach is suitable for mechanistic studies of complex multicomponent analyses involving membrane-bound systems.

show abstract

DNA-Directed Immobilization: Efficient, Reversible, and Site-Selective Surface Binding of Proteins by Means of Covalent DNA–Streptavidin Conjugates

Cited by 155 publications

References 26 publications

Biosensors and biochips: advances in biological and medical diagnostics

Biosensors and biochips: advances in biological and medical diagnostics

Microarray‐Based in vitro Evaluation of DNA Oligomer Libraries Designed in silico

Addressable adsorption of lipid vesicles and subsequent protein interaction studies

Contact Info

Product

Resources

About