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Self-assembly of DNA-streptavidin nanostructures and their use as reagents in immuno-PCR.

Abstract：

:The self-assembly of bis-biotinylated double-stranded DNA and the tetravalent biotin-binding protein streptavidin (STV) have been studied by non-denaturing gel electrophoresis and atomic force microscopy (AFM). The rapid self-assembly reproducibly generated populations of individual oligomeric complexes. Most strikingly, the oligomers predominantly contained bivalent STV molecules bridging two adjacent DNA fragments to form linear nanostructures. Trivalent STV branch points occurred with a lower frequency and the presence of tetravalent STV was scarce. However, valency distribution, size and the exchange dynamics of the supramolecular aggregates were highly sensitive to stoichiometric variations in the relative molar coupling ratio of bis-biotinylated DNA and STV. The largest aggregates were obtained from equimolar amounts while excess STV led to the formation of smaller oligomers appearing as fingerprint-like band patterns in electrophoresis. Excess DNA, however, induces a complete breakdown of the oligomers, likely a consequence of the instability of STV conjugates containing more than two biotinylated DNA fragments. It was demonstrated that the oligomers can further be functionalized, for instance by the coupling of biotinylated immunoglobulins. Both pure and also antibody-modified DNA-STV oligomers were used as reagents in immuno-PCR (IPCR), a highly sensitive detection method for proteins and other antigens. Employment of the supramolecular reagents led to an approximately 100-fold enhanced sensitivity compared to the conventional IPCR procedure.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Niemeyer CM,Adler M,Pignataro B,Lenhert S,Gao S,Chi L,Fuchs H,Blohm D

doi

10.1093/nar/27.23.4553

keywords:

subject

Has Abstract

pub_date

1999-12-01 00:00:00

pages

4553-61

issue

23

eissn

0305-1048

issn

1362-4962

pii

gkc664

journal_volume

27

pub_type

杂志文章

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