Maintenance of Nonviral Vector Particle Size During the Freezing Step of the Lyophilization Process is Insufficient for Preservation of Activity: Insight From Other Structural Indicators

Molina, Marion d.C.; Allison, Steven D.; Anchordoquy, Thomas J.

doi:10.1002/jps.1096

Cited by 78 publications

(64 citation statements)

References 38 publications

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“…Lyophilization is another method for addressing this problem and has been attested to be effective for long-term storage of nucleic acid formulations (7,13,14) and thus enables storage at room temperature. Besides cost reduction in transportation and storage, lyophilization is preferred over freezing due to better stability of biomolecules resulting from the removal of non freezable water associated with most biomolecules (15).…”

Section: Introductionmentioning

confidence: 99%

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Effect of Lyophilization and Freeze-thawing on the Stability of siRNA-liposome Complexes

et al. 2007

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Abstract. The purpose of this research was to describe the application of lyophilization in the delivery of siRNA using cationic lipids by addressing the long-term formulation/stability issues associated with cationic lipids and to understand the mechanism of lyoprotection. siRNA liposomes complexes were formed in different potential cyro/lyoprotectants and subjected to either lyophilization or freeze thaw cycles. siRNA, liposomes and/or lipoplexes were tested for activity, SYBR Green I binding, cellular uptake and particle size. The lipoplexes when lyophilized in the presence of sugars as lyoprotectants could be lyophilized and reconstituted without loss of transfection efficacy but in ionic solutions they lost 65-75% of their functionality. The mechanism of this loss of activity was further investigated. The lyophilization process did not alter siRNA's intrinsic biological activity as was evident by the ability of lyophilized siRNA to retain functionality and SYBR green I binding ability. While the lipoplex size dramatically increased (∼50-70 times) after lyophilization in the absence of non-ionic lyoprotectants. This increase in size correlated to the decrease in cellular accumulation of siRNA and a decrease in activity. In conclusion, siRNAs can be applied in cationic lipid lyophilized formulations and these complexes represent a potential method of increasing the stability of pre-formed complex.

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Section: Introductionmentioning

confidence: 99%

“…Dye accessibility assays are used to provide a measure of free siRNA not associated with the lipids and are based on the fact that SYBR Green I fluorescence is significantly increased upon binding to double stranded nucleic acids (14). Zipper et.…”

Section: Introductionmentioning

confidence: 99%

Effect of Lyophilization and Freeze-thawing on the Stability of siRNA-liposome Complexes

et al. 2007

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“…The storage conditions, especially for the long-term, should preserve any sterility treatment that was given to the product. Lyophilization is a technique that is widely used to store proteins/enzymes for long-term use [62][63][64] . It increases the shelf life of the lyophilized material by placing the product into a state of suspended activity due to dehydration.…”

Section: Discussionmentioning

confidence: 99%

Assessment of the Stability of an Immunoadsorbent for the Extracorporeal Removal of Beta-2-Microglobulin from Blood

2005

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Background: Dialysis-related amyloidosis (DRA) is a devastating and costly condition that affects patients with end stage kidney disease. A key feature of DRA is the formation of amyloid fibrils, consisting primarily of β₂-microglobulin. Except for kidney transplantation, conventional kidney replacement therapies, which are based on nonspecific mechanisms, do not adequately address β₂-microglobulin removal. An antihuman β₂-microglobulin single-chain variable region antibody fragment (scFv) was developed to confer specificity to β₂-microglobulin removal during hemodialysis. Methods: The scFv was immobilized onto agarose and characterized for β₂m binding capacity, thermal stability at 37°C, regeneration capacity, storage conditions, and sterility. Results: The β₂-microglobulin binding capacity was 1.3 mg/ml scFv gel. The immunoadsorbent is thermally stable, can be regenerated, stored short-term in 20% ethanol, lyophilized for long-term storage, and withstand process conditions similar to that of a patient’s hemodialysis therapy. Conclusions: The results support further investigation of immobilized scFvs as a novel tool to remove β₂-microglobulin from blood.

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“…63 Az egyik lehetséges megoldás a stabilitás kérdésére a részecske-szuszpenzó liofilizálása lehetne, azonban korábbi kísérletek azt mutatták, hogy a lineáris PEI-t tartalmazó formulációk nem liofilizálhatók (pontosabban liofilizálás után nem oldhatók vissza), annak ellenére, hogy az elágazó láncú PEI-vel ez a probléma nem jelentkezik. 64,65,66 Az AIDS halálos "betegségét" (Acquired Immune Deficiency Syndrome, vagyis szerzett immunhiányos tünet együttes) a HIV (Human Immunodeficiency Virus, vagyis emberi immunhiány vírus) okozza. 1983-ban a Pasteur Intézetben izoláltak egy vírust (többek között Françoise Barré-Sinoussi és Luc Montagnier), melyről úgy gondolták, hogy az AIDS-et okozza.…”

Section: 54unclassified

“…A PEI/pDNS rendszer e problémája már ismert az irodalomban, megoldásként pedig lioprotektorokat alkalmaznak, melyekkel javítható, de teljesen nem küszöbölhető ki a probléma, mert ha a visszaoldás sikeres is, a biológiai aktivitás mégsem éri el a liofilizálás előtti mértéket. 64,65 Lioprotektoroknak számítanak a poliolok, vagyis a hidroxid-csoportokban gazdag cukorszerű molekulák, mint például a trehalóz vagy a mannitol. A DermaVir szuszpenzió bár 7,5 tömeg% mannitolt tartalmaz a végösszetételben, ez nem bizonyult elég, vagy megfelelő lioprotektornak.…”

Section: A260unclassified

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Maintenance of Nonviral Vector Particle Size During the Freezing Step of the Lyophilization Process is Insufficient for Preservation of Activity: Insight From Other Structural Indicators

Cited by 78 publications

References 38 publications

Effect of Lyophilization and Freeze-thawing on the Stability of siRNA-liposome Complexes

Effect of Lyophilization and Freeze-thawing on the Stability of siRNA-liposome Complexes

Assessment of the Stability of an Immunoadsorbent for the Extracorporeal Removal of Beta-2-Microglobulin from Blood

Plazmid DNS-alapú nanomedicína formuláció fejlesztése

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