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Immobilization of Nucleic Acids Using Biotin-Strept(avidin) Systems

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Immobilisation of DNA on Chips II

Part of the book series: Topics in Current Chemistry ((TOPCURRCHEM,volume 261))

Abstract

There are several advantages for using biotin-streptavidin/avidin (strept(avi-din)) systems to immobilize nucleic acids and other molecules. These include the essential irreversible, but not covalent, binding of biotin to strept(avidin), the ease of biotinylating a large number of molecules without interfering with their function or the binding of biotin by strept(avidin), and the stability of strept(avidin) especially when bound with biotin. Another advantage of the biotin-strept(avidin) system is that it can be used for rapid prototyping to test a large number of protocols and molecules. The basic characteristics of the biotin-strept(avidin) are unique, although many of the approaches for immobilizing reagents with such systems are not unique. Here, biotin/strept(avidin) immobilizations systems are reviewed with an emphasis on nucleic acid applications.

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Abbreviations

BPL:

Biotin protein ligase

BSA:

Bovine serum albumin

Chimeric strept(avidin):

Multifunctional protein composed of a strept(avidin) portion and a second protein

Designer strept(avidin):

Strept(avidin) with increased functionality via recombinant protein modification

Dendrimer:

Regular, highly branched macromolecule with a monodisperse, tree-like or generational structure

DETA:

Trimethoxysilylpropyldiethylenetriamine

HMDS:

Hydrophobic hexamethyldisloxame

HMDS-PP:

Hydrophobic hexamethyldisloxame plasma polymer

Imino-biotin:

Guanido biotin derivative

Immuno-PCR:

Immuno-polymerase chain reaction detects the presence of an antigen by PCR amplification of a single-stranded DNA bound (directly or indirectly) to an antibody

K d :

Dissociation constant

MPTS:

(3-Mercaptopropyl)-trimethoxysilane

NHS:

N-Hydroxysuccinimide

NitroAvidin:

Example of chemical modified smart avidin that displays pH-dependent reversible binding of biotin

NutrAvidin:

Recombinant avidin proteins with a neutral charge created to reduce high backgrounds due to charge–charge interactions

PCR:

Polymerase chain reaction to amplify DNA exponentially

PEDA:

m,p(Amino-ethylamino-methyl)phenethyltrimethoxysilane

PDEAAm:

Poly(N,N-diethylacrylamide)

PNIPAAm:

Poly(N-isopropylacrylamide)

RCA:

Rolling circle amplification, an isothermal method of amplifying circular DNA using strand displacement

Smart strept(avidin)s:

A strept(avidin) that responds to its environment

SMPB:

Succinimidyl 4-[malemidophenyl]-butyrate

Strept(avidin):

Streptavidin and avidin

Supramolecular bioconjugates:

Created by hybridizing complementary single-stranded oligo nucleotide linked to different proteins (e.g., streptavidin and a second protein)

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Authors and Affiliations

  1. Molecular Biotechnology Research Laboratory, Boston University, 44 Cummington Street, 02215, Boston, MA, USA

    Cassandra L. Smith, Jaqueline S. Milea & Giang H. Nguyen

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  1. Cassandra L. Smith
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  2. Jaqueline S. Milea
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  3. Giang H. Nguyen
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Correspondence to Cassandra L. Smith .

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Christine Wittmann

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Smith, C.L., Milea, J.S., Nguyen, G.H. Immobilization of Nucleic Acids Using Biotin-Strept(avidin) Systems. In: Wittmann, C. (eds) Immobilisation of DNA on Chips II. Topics in Current Chemistry, vol 261. Springer, Berlin, Heidelberg. https://doi.org/10.1007/128_017

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