Freeze Drying Properties of Some Oligonucleotides

Jameel, Feroz; Amsberry, Kent L.; Pikal, Michael J.

doi:10.1081/pdt-100000743

Cited by 5 publications

(3 citation statements)

References 8 publications

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“…Furthermore the presence of pDNA had very little influence on the Tg´ (data not shown). This finding is in contrast to data shown by Jameel et al ., where oligonucleotides increase the Tg´ of lactose (30). Consequently, we lyophilised all formulations using the same lyophilisation cycle, resulting in a residual water content of approximately 0.8% ( w / w ) for the FD formulations (Table I).…”

Section: Resultsmentioning

confidence: 99%

Naked Plasmid DNA Formulation: Effect of Different Disaccharides on Stability after Lyophilisation

et al. 2010

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Since plasmid DNA (pDNA) is unstable in solution, lyophilisation can be used to increase product shelf life. To prevent stress on pDNA molecules during lyophilisation, cryo- and lyoprotectants have to be added to the formulation. This study assessed the effect of disaccharides on naked pDNA stability after lyophilisation using accelerated stability studies. Naked pDNA was lyophilised with sucrose, trehalose, maltose or lactose in an excipient/DNA w/w ratio of 20. To one part of the vials extra residual moisture was introduced by placing the vials half opened in a 25°C/60% RH climate chamber, before placing all vials in climate chambers (25°C/60% RH and 40°C/75% RH) for stability studies. An ex vivo human skin model was used to assess the effect of disaccharides on transfection efficiency. Lyophilisation resulted in amorphous cakes for all disaccharides with a residual water content of 0.8% w/w. Storage at 40°C/75% RH resulted in decreasing supercoiled (SC) purity levels (sucrose and trehalose maintained approximately 80% SC purity), but not in physical collapse. The addition of residual moisture (values between 7.5% and 10% w/w) resulted in rapid collapse except for trehalose and decreasing SC purity for all formulations. In a separate experiment disaccharide formulation solutions show a slight but significant reduction (<3% with sucrose and maltose) in transfection efficiency when compared to pDNA dissolved in water. We demonstrate that disaccharides, like sucrose and trehalose, are effective lyoprotectants for naked pDNA.

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Section: Resultsmentioning

confidence: 99%

Naked Plasmid DNA Formulation: Effect of Different Disaccharides on Stability after Lyophilisation

et al. 2010

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“…However, DNA must resist the freezing and drying stresses encountered during processing before stability can be addressed. In this respect, it has been demonstrated that the addition of disaccharides, such as sucrose and trehalose, are beneficial to prevent loss of SC DNA during lyophilization [55,56,82,91,96,113,115,121,178,179].…”

Section: Stability Of Dried Plasmid Dnamentioning

confidence: 99%

“…The interest in preserving DNA as dried preparations has increased vastly in response to the growing demand from a variety of fields, including pharmaceutical sciences [56,82,91,113,121,122], forensics [123], molecular diagnostics [124], and biorepository management [125]) for storing DNA samples or tumor specimen banks [114,126] for prolonged periods of time, sometimes in large numbers, in regions where sophisticated storage equipment may not be available. Although the requirements are quite different in each of these fields, the need for strategies that provide highly stable storage conditions (e.g., room temperature storage) seems to be a common factor.…”

Section: Preservation Of Dnamentioning

confidence: 99%