RESUMEN
Chromatographic methods have been essential tools for analysis and purification of synthetic oligonucleotides since the 1970s. Significant developments in terms of instruments and stationary phases (media) have been made during the past several decades; among the latest are sub-micron to micron particles for the media, as well as ultra performance liquid chromatography (UPLC). Micron and sub-micron particles have increased product resolution. Applications of recently developed methods such as IP-RP-HPLC and LC-MS have been discussed for analysis, along with use of various methods for purification. Utilization of UPLC has decreased analysis time, increasing the throughput for analysis. Commonly used methods for analysis and purification of synthetic oligonucleotides have been described in this unit.
Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Ácidos Nucleicos/química , Ácidos Nucleicos/aislamiento & purificaciónRESUMEN
A new class of non-explosive activators has been developed based on heterocyclic tertiary amine salts of saccharin. These salts have been found to be highly effective in the synthesis of oligonucleotides and nucleoside phosphoramidites.
Asunto(s)
Compuestos Heterocíclicos/química , Oligonucleótidos/síntesis química , Compuestos Organofosforados/síntesis química , Sacarina/análogos & derivados , Sustancias Explosivas/química , Compuestos Heterocíclicos/síntesis química , Sacarina/químicaRESUMEN
Significant developments in oligonucleotide chemistry, which occurred between 1980 and 2000, led to several oligonucleotide-based therapeutics entering clinical trials. Improvements in synthesis and purification methods, along with analytical techniques have facilitated the large-scale manufacture of high quality oligonucleotides to support these clinical trials.
Asunto(s)
Diseño de Fármacos , Oligonucleótidos/síntesis química , Compuestos Policíclicos/síntesis química , Técnicas de Química Analítica/métodos , Técnicas de Química Analítica/tendencias , Composición de Medicamentos/métodos , Composición de Medicamentos/tendencias , Humanos , Modelos Químicos , Estructura Molecular , Oligonucleótidos/química , Oligonucleótidos/uso terapéutico , Compuestos Policíclicos/química , Compuestos Policíclicos/uso terapéuticoRESUMEN
It is well known that exposure to UV induces DNA damage, which is the first step in mutagenesis and a major cause of skin cancer. Among a variety of photoproducts, cyclobutane-type pyrimidine photodimers (CPD) are the most abundant primary lesion. Despite its biological importance, the precise relationship between the structure and properties of DNA containing CPD has remained to be elucidated. Here, we report the free (unbound) crystal structure of duplex DNA containing a CPD lesion at a resolution of 2.0 A. Our crystal structure shows that the overall helical axis bends approximately 30 degrees toward the major groove and unwinds approximately 9 degrees, in remarkable agreement with some previous theoretical and experimental studies. There are also significant differences in local structure compared with standard B-DNA, including pinching of the minor groove at the 3' side of the CPD lesion, a severe change of the base pair parameter in the 5' side, and serious widening of both minor and major groves both 3' and 5' of the CPD. Overall, the structure of the damaged DNA differs from undamaged DNA to an extent that DNA repair proteins may recognize this conformation, and the various components of the replicational and transcriptional machinery may be interfered with due to the perturbed local and global structure.
