Large-scale synthesis oligonucleotides

PNA-oligomers are synthesized using methodology developed for peptide synthesis. The large-scale synthesis of oligonucleotides has been established for the preparation of sufficient quantities for clinical trials of antisense drugs. 

Wright P, Lloyd D, Rapp W, Andrus A. Large scale synthesis of oligonucleotides via phosphoramidite nucleosides and a high loaded polystyrene support. Tetrahedron Lett 34 3373-3376, 1993. 

Bardella F, Giralt E, Pedroso E. Polystyrene supported synthesis by the phosphite triester approach An alternative for the large scale synthesis of small oligonucleotides. Tetrahedron Lett 31 6231-6234, 1990. 

Bonora, G M, Biancotto, G, and Scremin, C L (1991) Use of the amidite chemistry in the PEG-supported, large-scale synthesis of oligonucleotides The HELP Plus method Nucl Acids Symp Ser 24, 222. 

In order to achieve good-quality synthesis on a large scale, proper setting of the instrument is an integral part of the process. Here, large-scale synthesis of oligonucleotides on solid-phase has been discussed using a model 8800 DNA synthesizer. The instrument 8800 DNA synthesizer consists of different components ... 

Prior to 1990, this step was commonly performed with an elemental sulfur solution in a pyridine-carbondisulfide mixture. The utilization of [3H]-1,2-benzodithiol-3-one-1,1 -dioxide by Iyer et al. (15) for converting phosphorus (III) triester into pentavalent phosphorothioate has simplified the process of large-scale polyphosphorothioated oligonucleotide synthesis following phosphoramidite chemistry. This reagent, however, fails to convert H-phosphonate diester linkages into phosphorothioate diester, which is still achieved by treating with 5% elemental sulfur solution in pyridine-carbon disulfide off the instrument. 

Wright, P., Lyttle, M., Carrol, J, Hudson, D, Warren, W., and Sinha, N. D, Large scale synthesis of oligodeoxynucleotides and polyphosphorothioated oligonucleotides, presented at the International Conference on Nucleic Acid Therapeutics held on January 13-17, 1991 in Clearwater Beach, Florida. 

Synthetic organic polymers, which are used as polymeric supports for chromatography, as catalysts, as solid-phase supports for peptide and oligonucleotide synthesis, and for diagnosis, are based mainly on polystyrene, polystyrene-divinylbenzene, polyacrylamide, polymethacrylates, and polyvinyl alcohols. A conventional suspension of polymerization is usually used to produce these organic polymeric supports, especially in large-scale industrial production. 

Nucleotides and nucleic acids are critical tools in the areas of gene expression, therapeutics, and diagnostics. However, there are certain challenges associated with their large-scale purification and subsequent characterization. While solid-state oligonucleotide synthesis is relatively simple and can be totally automated, intra- and intermolecular associations may occur involving shorter sequences that may hybridize with the desired full length... 

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