Conjugation of nucleic acid probes to 6-aminoglucose-based homo- and copolymers. II. Application to diagnostics

Delair, Thierry; Badey, Bérengère; Charles, Marie‐Hélène; Laayoun, Ali; Domard, Alain; Pichot, Christian; Mandrand, Bernard

doi:10.1002/(sici)1099-1581(199709)8:9<545::aid-pat685>3.0.co;2-s

Cited by 3 publications

(2 citation statements)

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“…The second step is the reaction of the ODN derivative with the activated ester function (Scheme 1 ). As observed in earlier works ( 25 , 26 ), determining optimum experimental conditions called for carrying out numerous preliminary tests in different aqueous and dimethylformamide (DMF)/H 2 O buffers, at various ionic strengths (0.1–0.4 M NaCl), and for various ODN/A8–35 ratios and crosslinker concentrations (see the Materials and Methods section).…”

Section: Resultsmentioning

confidence: 99%

Synthesis of an oligonucleotide-derivatized amphipol and its use to trap and immobilize membrane proteins

Bon

Pia

Giusti

et al. 2014

Nucleic Acids Research

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Amphipols (APols) are specially designed amphipathic polymers that stabilize membrane proteins (MPs) in aqueous solutions in the absence of detergent. A8–35, a polyacrylate-based APol, has been grafted with an oligodeoxynucleotide (ODN). The synthesis, purification and properties of the resulting ‘OligAPol’ have been investigated. Grafting was performed by reacting an ODN carrying an amine-terminated arm with the carboxylates of A8–35. The use of OligAPol for trapping MPs and immobilizing them onto solid supports was tested using bacteriorhodopsin (BR) and the transmembrane domain of Escherichia coli outer membrane protein A (tOmpA) as model proteins. BR and OligAPol form water-soluble complexes in which BR remains in its native conformation. Hybridization of the ODN arm with a complementary ODN was not hindered by the assembly of OligAPol into particles, nor by its association with BR. BR/OligAPol and tOmpA/OligAPol complexes could be immobilized onto either magnetic beads or gold nanoparticles grafted with the complementary ODN, as shown by spectroscopic measurements, fluorescence microscopy and the binding of anti-BR and anti-tOmpA antibodies. OligAPols provide a novel, highly versatile approach to tagging MPs, without modifying them chemically nor genetically, for specific, reversible and targetable immobilization, e.g. for nanoscale applications.

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Section: Resultsmentioning

confidence: 99%

Synthesis of an oligonucleotide-derivatized amphipol and its use to trap and immobilize membrane proteins

Bon

Pia

Giusti

et al. 2014

Nucleic Acids Research

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“…Among the many synthesis routes that have been explored are the following: (i) formation of an amide bond between an ODN functionalized with a primary amine and appropriate functions carried by the polymer, such as a carboxylic acid, an anhydride or an NHS group (Ferraton et al 1997; Jeong et al 2005); (ii) creation of a disulfide bridge between thiols carried by the polymer and by a chemically modified ODN (Oishi et al 2005); (iii) Michaël addition between a maleimide function carried by the polymer and a thiolated ODN; and (iv) reductive amination, with the formation of an alkylamine, between an aldehyde-carrying polymer and the amine function of a modified ODN (Delair et al 1997; Ferraton et al 1997). …”

Section: Labeled and Functionalized Versions Of A8-35mentioning

confidence: 99%

Labeling and Functionalizing Amphipols for Biological Applications

2014

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Amphipols (APols) are short amphipathic polymers developed as an alternative to detergents for handling membrane proteins (MPs) in aqueous solution. MPs are, as a rule, much more stable following trapping with APols than they are in detergent solutions. The best-characterized APol to date, called A8-35, is a mixture of short-chain sodium polyacrylates randomly derivatized with octylamine and isopropylamine. Its solution properties have been studied in detail, and it has been used extensively for biochemical and biophysical studies of MPs. One of the attractive characteristics of APols is that it is relatively easy to label them, isotopically or otherwise, without affecting their physical-chemical properties. Furthermore, several variously modified APols can be mixed, achieving multiple functionalization of MP/APol complexes in the easiest possible manner. Labeled or tagged APols are being used to study the solution properties of APols, their miscibility, their biodistribution upon injection into living organisms, their association with MPs and the composition, structure and dynamics of MP/APol complexes, examining the exchange of surfactants at the surface of MPs, labeling MPs to follow their distribution in fractionation experiments or to immobilize them, increasing the contrast between APols and solvent or MPs in biophysical experiments, improving NMR spectra, etc. Labeling or functionalization of APols can take various courses, each of which has its specific constraints and advantages regarding both synthesis and purification. The present review offers an overview of the various derivatives of A8-35 and its congeners that have been developed in our laboratory and discusses the pros and cons of various synthetic routes.

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Nucleic Acid Analysis

Christopoulos

1999

Anal. Chem.

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The rapid progress in the Human Genome Project has stimulated the development of diverse analytical methods for mutation detection, elucidation of complex biological problems, molecular diagnosis and prognosis of disease, and assessment of treatment. This review will discuss recent advances in nucleic acid analysis as reported in articles that appeared in Chemical Abstracts between October 1, 1996 and October 1, 1998. Major areas of nucleic acid analysis covered in this review include purification methods, hybridization, in situ hybridization, DNA sequencing, amplification techniques, oligonucleotide arrays, miniaturization of nucleic acid analysis, and nucleic acid sensors.

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Conjugation of nucleic acid probes to 6-aminoglucose-based homo- and copolymers. II. Application to diagnostics

Cited by 3 publications

References 6 publications

Synthesis of an oligonucleotide-derivatized amphipol and its use to trap and immobilize membrane proteins

Synthesis of an oligonucleotide-derivatized amphipol and its use to trap and immobilize membrane proteins

Labeling and Functionalizing Amphipols for Biological Applications

Nucleic Acid Analysis

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