title: Lipid nanoparticle purification by spin centrifugation-dialysis (SCD): a facile and high-throughput approach for small scale preparation of siRNA-lipid complexes.
abstract: This paper describes the use of spin centrifugation-dialysis (SCD) for small-scale concentration/purification of siRNA-lipid complexes designed for use as therapeutic agents for gene silencing. SCD consists of a two-step method for concentration, filtration and buffer exchange of lipid nanoparticles (LNP) to provide a homogeneous preparation suitable for injection. Here, we compare SCD with the more traditionally used tangential flow filtration (TFF), and demonstrate the physicochemical and biological comparability of LNPs produced with both methods. TFF is a highly scalable method used in both developmental and production applications, but is limited in terms of miniaturization. In contrast to TFF, SCD is faster, less expensive, and requires less oversight for assembling LNPs for small-scale applications, such as target screening both in vitro and in vivo. The finding that SCD is a viable method for filtering LNPs in a manner similar to TFF, producing particles with comparable properties and biological activity, is significant given the complexity and sensitivity of LNPs to processing conditions.
authors: Sirna Therapeutics, wholly owned subsidiary of Merck and Co., 1700 Owens Street, Fourth Floor, San Francisco, CA 94158, USA.
authors: Radu
authors: R
authors: Mihaila
authors: Silvia
authors: S
authors: Chang
authors: Alan T
authors: AT
authors: Wei
authors: Zhi Y
authors: ZY
authors: Hu
authors: Dipali
authors: D
authors: Ruhela
authors: Timothy R
authors: TR
authors: Shadel
authors: Sven
authors: S
authors: Duenwald
authors: Elmer
authors: E
authors: Payson
authors: James J
authors: JJ
authors: Cunningham
authors: Nelly
authors: N
authors: Kuklin
authors: David J
authors: DJ
authors: Mathre
mesh_terms: D000818:Animals; D002021:Buffers; D045744:Cell Line, Tumor; D002498:Centrifugation / Q000295:instrumentation; D003956:Dialysis / Q000295:instrumentation / Q000379:methods*; D015536:Down-Regulation; D004867:Equipment Design; D005374:Filtration; D057166:High-Throughput Screening Assays / Q000295:instrumentation; D006863:Hydrogen-Ion Concentration; D008055:Lipids / Q000737:chemistry*; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D008904:Miniaturization; D053758:Nanoparticles; D036103:Nanotechnology; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000737:chemistry* / Q000378:metabolism; D013997:Time Factors; D014162:Transfection
chemical_list: D002021:Buffers; D008055:Lipids; D034741:RNA, Small Interfering; D008156:Luciferases