A fast and sensitive method for measuring the integrity of siRNA-carrier complexes in full human serum.
SiRNA based therapeutics are currently under investigation for treatment of cancer and viral infections. Upon intravenous administration, the nanoscopic delivery systems which carry the siRNA need to be stable in serum, an aspect which is often overlooked in numerous publications on siRNA delivery systems. Techniques currently available for studying the dissociation of siRNA-liposome complexes are time consuming and incompatible with full serum. We therefore developed a fluorescence fluctuation spectroscopy (FFS) based method which allows to monitor the integrity of siRNA-carrier complexes. The method can very rapidly provide quantitative information on the complex integrity in biological media, like full human serum, and at very low siRNA concentrations (approximately 20 nM siRNA). Information on the integrity of intravenously injected siRNA nanoparticles in serum is crucial. Consequently, the FFS method reported in this work may find broad applicability in the field of siRNA-carrier design.
Buyens
Kevin
K
Laboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmacy, Ghent University, Harelbekestraat 72, 9000 Ghent, Belgium.
Lucas
Bart
B
Raemdonck
Koen
K
Braeckmans
Kevin
K
Vercammen
Jo
J
Hendrix
Jelle
J
Engelborghs
Yves
Y
De Smedt
Stefaan C
SC
Sanders
Niek N
NN
D004337:Drug Carriers / Q000737:chemistry*; D004355:Drug Stability; D004591:Electrophoresis, Polyacrylamide Gel; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry; D006801:Humans; D007700:Kinetics; D008081:Liposomes; D010714:Phosphatidylethanolamines / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D034741:RNA, Small Interfering / Q000097:blood* / Q000737:chemistry; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence; D013997:Time Factors
C410262:1,2-dioleoyl-glycero-3-phosphatidyl ethanolamine; C519184:1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-methoxy-poly(ethylene glycol 2000); D004337:Drug Carriers; D005229:Fatty Acids, Monounsaturated; D008081:Liposomes; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; D034741:RNA, Small Interfering; D011092:Polyethylene Glycols; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane