Oligodeoxyribonucleotide Activation with 2,4-Phenylenediisothiocyanate and Their Covalent Grafting onto Amine-Functionalized Latex Microspheres

The activation of various synthetic oligodeoxyribonucleotides (ODNs), bearing a hexamethylamine arm at their 5′-end, with the 2,4-phenylenediisothiocyanate (PDC) was studied. The activation reaction had to be performed within 40 min, because of competitive hydrolysis of the isothiocyanate functions....

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Bibliographic Details
Published inJournal of dispersion science and technology Vol. 22; no. 5; pp. 473 - 484
Main Authors Ganachaud, F., Laayoun, A., Chaix, C., Delair, Th, Pichot, C., Elaïssari, A.
Format Journal Article
LanguageEnglish
Published Philadelphia, PA Taylor & Francis Group 31.07.2001
Taylor & Francis
Subjects
2,4-Phenylene diisothiocyanate
Activation analysis
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Chemical Sciences
Electrophoresis
Enzymes
Fundamental and applied biological sciences. Psychology
General and theoretical biochemistry
Grafting (chemical)
HPLC
Hydrolysis
Ionization
Latex
Life Sciences
MALDI-TOF
Mass spectrometers
Molecular and cellular biology
Nucleic acid sequences
Oligodeoxyribonucleotides
Pharmaceutical sciences
Surface modification
Oligodeoxyribonucleotide
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Summary:The activation of various synthetic oligodeoxyribonucleotides (ODNs), bearing a hexamethylamine arm at their 5′-end, with the 2,4-phenylenediisothiocyanate (PDC) was studied. The activation reaction had to be performed within 40 min, because of competitive hydrolysis of the isothiocyanate functions. ODN modifications were then characterized by different analytical methods i.e. gel capillary electrophoresis, Matrix-Assisted Laser Desorption-Ionization-Time of Flight (MALDI-TOF) mass spectrometry. Moreover, activation of the amine groups of the nucleic bases was evidenced through the chemical analysis of the enzymatic-digested conjugate (PDC-ODN) by reverse phase HPLC. Preliminary results on the covalent grafting of activated ODN molecules onto the aminated latex particles showed that the amount of grafted ODNs was directly related to the number of potential grafting sites on the nucleic acid probe (primary amine group of hexamethyl arm and amine functions of the nucleic bases).
Bibliography:ObjectType-Article-2
SourceType-Scholarly Journals-1
ObjectType-Feature-1
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ISSN:0193-2691
1532-2351
DOI:10.1081/DIS-100107856