A flexible, thermostable nanostructured lipid carrier platform for RNA vaccine delivery

Current RNA vaccines against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) are limited by instability of both the RNA and the lipid nanoparticle delivery system, requiring storage at −20°C or −70°C and compromising universally accessible vaccine distribution. This study demonstrates the thermostability and adaptability of a nanostructured lipid carrier (NLC) delivery system for RNA vaccines that has the potential to address these concerns. Liquid NLC alone is stable at refrigerated temperatures for ≥1 year, enabling stockpiling and rapid deployment by point-of-care mixing with any vaccine RNA. Alternatively, NLC complexed with RNA may be readily lyophilized and stored at room temperature for ≥8 months or refrigerated temperature for ≥21 months while still retaining the ability to express protein in vivo. The thermostability of this NLC/RNA vaccine delivery platform could significantly improve distribution of current and future pandemic response vaccines, particularly in low-resource settings. Current RNA vaccines against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) are limited by instability of both the RNA and the lipid nanoparticle delivery system, requiring storage at −20°C or −70°C and compromising universally accessible vaccine distribution. This study demonstrates the thermostability and adaptability of a nanostructured lipid carrier (NLC) delivery system for RNA vaccines that has the potential to address these concerns. Liquid NLC alone is stable at refrigerated temperatures for ≥1 year, enabling stockpiling and rapid deployment by point-of-care mixing with any vaccine RNA. Alternatively, NLC complexed with RNA may be readily lyophilized and stored at room temperature for ≥8 months or refrigerated temperature for ≥21 months while still retaining the ability to express protein in vivo. The thermostability of this NLC/RNA vaccine delivery platform could significantly improve distribution of current and future pandemic response vaccines, particularly in low-resource settings. IntroductionRNA-based vaccines show great promise to effectively address existing and emerging infectious diseases,1Deering R.P. Kommareddy S. Ulmer J.B. Brito L.A. Geall A.J. Nucleic acid vaccines: prospects for non-viral delivery of mRNA vaccines.Expert Opin. Drug Deliv. 2014; 11: 885-899Crossref PubMed Scopus (105) Google Scholar, 2Rauch S. Jasny E. Schmidt K.E. Petsch B. New vaccine technologies to combat outbreak situations.Front Immunol. 2018; 9: 1963Crossref PubMed Scopus (298) Google Scholar, 3Zhang C. Maruggi G. Shan H. Li J. Advances in mRNA vaccines for infectious diseases.Front Immunol. 2019; 10: 594Crossref PubMed Scopus (322) Google Scholar including the ongoing pandemic caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). RNA vaccines can be rapidly adapted to new targets and manufactured using sequence-independent operations, thus reducing the cost and time to develop new vaccines, particularly in pandemic settings.4Pardi N. Hogan M.J. Porter F.W. Weissman D. mRNA vaccines a new era in vaccinology.Nat. Rev. Drug Discov. 2018; 17: 261-279Crossref PubMed Scopus (1516) Google Scholar The Emergency Use Authorization granted to two safe and highly effective mRNA vaccines targeting SARS-CoV-2, less than 1 year after sequencing the novel coronavirus, highlights the power of this new technology.5Jackson L.A. Anderson E.J. Rouphael N.G. Roberts P.C. Makhene M. Coler R.N. McCullough M.P. Chappell J.D. Denison M.R. Stevens L.J. et al.An mRNA vaccine against SARS-CoV-2 - preliminary report.N. Engl. J. Med. 2020; 383: 1920-1931Crossref PubMed Scopus (1675) Google Scholar,6Polack F.P. Thomas S.J. Kitchin N. Absalon J. Gurtman A. Lockhart S. Perez J.L. Perez Marc G. Moreira E.D. Zerbini C. et al.Safety and efficacy of the BNT162b2 mRNA covid-19 vaccine.N. Engl. J. Med. 2020; 383: 2603-2615Crossref PubMed Scopus (6139) Google Scholar However, one of the biggest challenges facing these extraordinary new vaccines is the ability to successfully distribute them widely in the face of a pandemic. Cold chain storage is required for both authorized vaccines (−70°C and −20°C for the SARS-CoV-2 RNA vaccines produced by Pfizer/BioNTech and Moderna, respectively). Frozen shipping and storage even at standard freezer conditions poses difficulties in settings with well-established medical infrastructure, challenges greatly compounded in areas with limited resources.7Kumru O.S. Joshi S.B. Smith D.E. Middaugh C.R. Prusik T. Volkin D.B. Vaccine instability in the cold chain: mechanisms, analysis and formulation strategies.Biologicals. 2014; 42: 237-259Crossref PubMed Scopus (208) Google Scholar, 8Chen D. Zehrung D. Desirable attributes of vaccines for deployment in low-resource settings.J. PubMed Scopus Google Scholar, Volkin D.B. E. the cold of mRNA vaccine PubMed Scopus Google of in RNA vaccines is a the this are and Volkin D.B. E. the cold of mRNA vaccine PubMed Scopus Google Scholar, D. G. nanoparticle vaccines: and J. PubMed Scopus Google Scholar, of storage and of covid-19 9: PubMed Scopus Google Scholar However, are vaccine RNA are to by of the RNA has in to by and a new of Rev. Drug Discov. 2014; PubMed Scopus Google Scholar to and RNA alone a to Anderson in Rev. Drug Discov. PubMed Scopus Google Scholar RNA delivery are to and RNA by et A. Anderson the in technologies for mRNA 2019; PubMed Scopus Google Scholar and et S. J. in delivery of mRNA using delivery PubMed Scopus Google The current system of for RNA vaccines, including SARS-CoV-2 vaccines in to is a lipid nanoparticle delivery L.A. Anderson E.J. Rouphael N.G. Roberts P.C. Makhene M. Coler R.N. McCullough M.P. Chappell J.D. Denison M.R. Stevens L.J. et al.An mRNA vaccine against SARS-CoV-2 - preliminary report.N. Engl. J. Med. 2020; 383: 1920-1931Crossref PubMed Scopus (1675) Google S. Advances in lipid for PubMed Scopus Google Scholar, for 2014; PubMed Scopus Google Scholar, A. D. mRNA vaccine delivery using lipid Deliv. PubMed Scopus Google Scholar, A. Brito L.A. Smith M. J. et and of by mRNA vaccines against and PubMed Scopus Google Scholar in the RNA is a lipid This in that the RNA and by the A. D. mRNA vaccine delivery using lipid Deliv. PubMed Scopus Google K.E. E. A. A. S. J. T. et of lipid for of mRNA Nucleic 2019; PubMed Scopus Google Scholar However, of both the RNA and Volkin D.B. E. the cold of mRNA vaccine PubMed Scopus Google Scholar, D. G. nanoparticle vaccines: and J. PubMed Scopus Google Scholar, of storage and of covid-19 9: PubMed Scopus Google Scholar with to temperatures in to after of lipid the of and J. PubMed Scopus Google J. S. Li G. storage of for mRNA 2020; PubMed Scopus Google Scholar of to the thermostability of RNA vaccines at A. M. D. S. T. T. T. Petsch B. RNA vaccine against 11: PubMed Scopus Google Scholar, Li H. G. C. et mRNA vaccine against 2020; PubMed Scopus Google Scholar, H. Li Li C. G. Li et and efficacy of the mRNA vaccine in PubMed Scopus Google Scholar authorized RNA vaccines in the still Vaccine - - of delivery and to RNA L.A. M. A. T. M. J. A. et for the delivery of RNA 2014; PubMed Scopus Google Scholar, J. B. J. M. E. J. E. S. et nanostructured lipid carrier for delivery of a RNA against 2018; PubMed Scopus Google Scholar, of lipid nanoparticle for and in delivery of 2019; PubMed Scopus Google Scholar the ability of a nanostructured lipid carrier (NLC) system to effectively vaccines by This NLC delivery system can be complexed and lyophilized with The NLC alone for at 1 year of storage at refrigerated while lyophilized NLC/RNA to and ability to protein in after at months of room temperature storage and at months at refrigerated of NLC delivery system a NLC delivery system by et J. B. J. M. E. J. E. S. et nanostructured lipid carrier for delivery of a RNA against 2018; PubMed Scopus Google of of and by and and a lipid This system is in the of RNA. The NLC system in the of a at and for at 1 year and a vaccine RNA is with the and the NLC/RNA the in the RNA and the in the of the NLC this the RNA at the of the NLC a RNA the protein the NLC is to ability to with the RNA after storage of the NLC for at months at refrigerated temperatures The complexed RNA is by the RNA is This is with of RNA complexed the of by of lipid nanoparticle for and in delivery of 2019; PubMed Scopus Google Scholar this is even after storage of the NLC a to to this NLC is for stockpiling for pandemic RNA targeting a can be rapidly produced in response to a pandemic and complexed with and of the of vaccine against that of and against J. B. J. M. E. J. E. S. et nanostructured lipid carrier for delivery of a RNA against 2018; PubMed Scopus Google Scholar that the of this vaccine can be successfully lyophilized for potential storage with the of a The of the of a lyophilized and to the of the system against the and of lyophilized vaccine with complexed of or conditions after has the NLC and of after has with and the NLC The and vaccines for of RNA and after of storage at analysis of the is in of and vaccine by at in in of and vaccine by the standard and is after by of RNA NLC/RNA and both the complexed and the vaccines are stable for at at refrigerated temperatures and retaining ability to the RNA with both and lyophilized and the lyophilized vaccine is to to vaccine at the 1 that the and of this The of the and this to in the of NLC for NLC/RNA vaccines are readily has the potential to significantly the challenges of RNA vaccines in both pandemic and of vaccines the in the ability of a delivery system to effectively vaccine mRNA is a and in the The and of this system is by with mRNA of the of the mRNA against by and with the the of the to the in that with the vaccine with in a and that is to that in that are and This demonstrates of for of in to of of lyophilized or with complexed of mRNA or lyophilized conditions after has the NLC and of mRNA after has with and the NLC analysis of the is in of and lyophilized by the standard of lyophilized the thermostability of the RNA vaccine platform using a RNA system of with a stored at and are with stored at and stored at and and analysis lyophilized the study with or and lyophilized readily with the for the NLC/RNA or storage conditions in with complexed of and at of the time with a complexed the standard RNA of the stored by at and and after with at time at time in and analysis is in in for or stored in with complexed after of to the of the at time the standard in at months for lyophilized vaccine stored at vaccine stored at and the a and in including and lyophilized of less than are the and time for conditions for stored at This demonstrates the of NLC/RNA them to the of the and even at temperatures for lyophilized and for is to while is for stored at this the ability of the to protein in and in the is after and after by after of the RNA the stored NLC and this is after storage and with show with the complexed for stored lyophilized at for and for stored lyophilized at and at and −20°C to months these storage conditions after with the RNA is still is RNA is after for stored lyophilized at or at and after months of of the RNA after is at for the at for the and at months for the lyophilized and ability of stored to express protein in is by of of by of and analysis of by time a a and a that of a a at time to this in protein in with the RNA by to the complexed in the of and of a RNA after in and of conditions is at for the at for the and at months for the lyophilized with the by months of stored lyophilized at and and at and −20°C in the time the lyophilized and show a in to the complexed still significantly for both conditions is after months of in the of in for the lyophilized and with the complexed the of these stored at months with the complexed the in at time in is to a in a than a of the of the stored This is by the in in the complexed analysis with the stored the ability of these to in after storage demonstrates the potential for thermostability of this delivery vaccines are to combat existing and emerging infectious including SARS-CoV-2, to rapid adaptability to new R.P. Kommareddy S. Ulmer J.B. Brito L.A. Geall A.J. Nucleic acid vaccines: prospects for non-viral delivery of mRNA vaccines.Expert Opin. Drug Deliv. 2014; 11: 885-899Crossref PubMed Scopus (105) Google Scholar, 2Rauch S. Jasny E. Schmidt K.E. Petsch B. New vaccine technologies to combat outbreak situations.Front Immunol. 2018; 9: 1963Crossref PubMed Scopus (298) Google Scholar, 3Zhang C. Maruggi G. Shan H. Li J. Advances in mRNA vaccines for infectious diseases.Front Immunol. 2019; 10: 594Crossref PubMed Scopus (322) Google Scholar, N. Hogan M.J. Porter F.W. Weissman D. mRNA vaccines a new era in vaccinology.Nat. Rev. Drug Discov. 2018; 17: 261-279Crossref PubMed Scopus (1516) Google Scholar, L.A. Anderson E.J. Rouphael N.G. Roberts P.C. Makhene M. Coler R.N. McCullough M.P. Chappell J.D. Denison M.R. Stevens L.J. et al.An mRNA vaccine against SARS-CoV-2 - preliminary report.N. Engl. J. Med. 2020; 383: 1920-1931Crossref PubMed Scopus (1675) Google Scholar, F.P. Thomas S.J. Kitchin N. Absalon J. Gurtman A. Lockhart S. Perez J.L. Perez Marc G. Moreira E.D. Zerbini C. et al.Safety and efficacy of the BNT162b2 mRNA covid-19 vaccine.N. Engl. J. Med. 2020; 383: 2603-2615Crossref PubMed Scopus (6139) Google Scholar However, cold chain for current RNA vaccine greatly distribution and to for rapid in the of RNA vaccine Volkin D.B. E. the cold of mRNA vaccine PubMed Scopus Google D. G. nanoparticle vaccines: and J. PubMed Scopus Google Scholar that a safe and effective RNA vaccine delivery J. B. J. M. E. J. E. S. et nanostructured lipid carrier for delivery of a RNA against 2018; PubMed Scopus Google Scholar has potential to greatly thermostability to current The NLC alone is stable at refrigerated temperatures for than 1 NLC complexed with mRNA or is to be lyophilized with both lyophilized and of after storage for of RNA vaccine by for at of storage at refrigerated This delivery may for RNA vaccine and cost to the thermostability of delivery to the of the NLC in of and of for at 1 year refrigerated refrigerated storage for the SARS-CoV-2 vaccines is limited in the storage of for to 1 at Vaccine - Scholar and Pfizer/BioNTech to of - Scholar that may be stable for months at refrigerated temperatures with of lipid the of and J. PubMed Scopus Google H. with storage in PubMed Scopus Google Scholar the limited of vaccines at refrigerated temperatures is to limited mRNA even with the RNA in the delivery than instability of the delivery D. G. nanoparticle vaccines: and J. PubMed Scopus Google the NLC system to RNA against authorized RNA vaccines the RNA in the of the However, this is to and RNA to of lipid nanoparticle for and in delivery of 2019; PubMed Scopus Google Scholar the NLC delivery system, the and the of the NLC/RNA and of the RNA by storage and after with and the of this RNA vaccine formulation a to vaccines and and a cold chain O.S. Joshi S.B. Smith D.E. Middaugh C.R. Prusik T. Volkin D.B. Vaccine instability in the cold chain: mechanisms, analysis and formulation strategies.Biologicals. 2014; 42: 237-259Crossref PubMed Scopus (208) Google D. Zehrung D. Desirable attributes of vaccines for deployment in low-resource settings.J. PubMed Scopus Google S. B. and the of for PubMed Scopus Google Scholar, D. E.J. storage of RNA for in vaccine PubMed Scopus Google Scholar, B. M. E. B. D. T. A. et efficacy of in mRNA vaccines against PubMed Scopus Google Scholar, M. G. A. et al.Safety and of a mRNA vaccine in 1 PubMed Scopus Google Scholar lyophilized mechanisms, in with the of the system or the system the of the or is S. E. of still still 2018; 10: Scopus Google Scholar mRNA to be the in the of current address that RNA alone to be to with et D. E.J. storage of RNA for in vaccine PubMed Scopus Google Scholar that lyophilized RNA ability to protein after storage at for to However, of has for by et S. E. of still still 2018; 10: Scopus Google Scholar and et and current Drug Deliv. Rev. 2019; PubMed Scopus Google can be to the a lipid a The and of may in S. E. of still still 2018; 10: Scopus Google and current Drug Deliv. Rev. 2019; PubMed Scopus Google Scholar the of is still J. A. A.J. S. of lipid or mRNA by B. 2018; PubMed Scopus Google Scholar, M. T. S. S. A.J. A. N. A. J. et of protein mRNA by lipid A. 2018; PubMed Scopus Google Scholar, S. E. M.J. by mixing nanostructured PubMed Scopus Google Scholar and may and D. G. nanoparticle vaccines: and J. PubMed Scopus Google Scholar may still be to the and instability at vaccine after of lipid the of and J. PubMed Scopus Google Scholar or of RNA after storage even with the of J. S. Li G. storage of for mRNA 2020; PubMed Scopus Google Scholar of may be by and C. D. C. of and future PubMed Scopus Google the of and RNA vaccine delivery is and the of the NLC delivery system is to than to are a with this system the RNA is complexed to the of the and of RNA and in lyophilized vaccines potential for vaccine N. et of a vaccine against using a J. 2018; PubMed Scopus Google potential of the NLC delivery system for pandemic response is and This and to the in vaccines, to pandemic The NLC system the of to response is the with E. C. M. et the efficacy of mRNA and protein vaccines by and PubMed Scopus Google Scholar the system the of with RNA produced by the NLC can the lipid the of in to authorized vaccines, the NLC delivery system is manufactured the RNA. pandemic the NLC alone could be to rapid is manufactured RNA of or with may be rapidly and complexed the of the rapid vaccine to or emerging that the of the lyophilized NLC system is with RNA a protein than a vaccine of a protein is a of lipid the of and J. PubMed Scopus Google of lipid nanoparticle for and in delivery of 2019; PubMed Scopus Google S. J. E. B. mRNA with a of in Nucleic PubMed Scopus Google Scholar a of with vaccine including of vaccine is a for of this However, is that vaccine be to that of the protein of the NLC formulation alone is for to any vaccine while this formulation has are and are in and Drug is in the and is to in and the lipid has successfully in the current study demonstrates the thermostability of NLC/RNA this a for this system and of the formulation or or be to this system to the of and conditions and to with vaccine of a RNA for could the of a system in of in this while potential that can of the NLC/RNA formulation may to significantly and distribution. vaccine in the face of a pandemic and the NLC delivery system for RNA vaccine delivery to improve vaccine distribution both and in the and for the and produced J. B. J. M. E. J. E. S. et nanostructured lipid carrier for delivery of a RNA against 2018; PubMed Scopus Google Scholar for the at the of the protein the and for and a The the and the of with the the of a the vaccine J. B. J. M. E. J. E. S. et nanostructured lipid carrier for delivery of a RNA against 2018; PubMed Scopus Google Scholar with a of the to to to current and of the for for RNA the protein in two and The is to that by et J. B. J. M. E. J. E. S. et nanostructured lipid carrier for delivery of a RNA against 2018; PubMed Scopus Google Scholar and the for in the in the and to with in the vaccine in using in and using or and by by and of by in using using using and and to the by and RNA the and by using a by and using The with a and stored at by and both by and mRNA a formulation NLC formulation J. B. J. M. E. J. E. S. et nanostructured lipid carrier for delivery of a RNA against 2018; PubMed Scopus Google Scholar and and at in a in and to in a the and at in a The by to the at for The NLC with a and stored at formulation of and in the NLC by in in of of at using system in with a The a with a system of a of and a of with both and The system at and at a of 1 and in and the to a standard at a of for by mixing RNA by with NLC in a and to a RNA in and for with or in the for after mixing to using a by the The of a at a at and and a at and the of the to with and to the using and to and to any in of both the NLC formulation alone and the NLC/RNA using in in and in a with the of RNA after and against by by or to a RNA of in the RNA with for at room temperature at to RNA of and This by with at a of for at both and RNA the by to the by and at for The for by with and at for of RNA and a at for in RNA and conditions in a RNA with at and using a system analysis of the using and RNA with the of RNA or a complexed for of RNA in of RNA in in the and of in to a of and in to a of The by to alone or in for to and and at for to the and the the and the the for both the RNA and the RNA the by with in for to a of for and in to the and to using the and and of at study The for in this the of the and Use the and Use in with of the of the and and the for the and Use of in of complexed vaccines, with 1 of complexed 1 of or of a Vaccine in in both of for a of of vaccine for of for the with by the for by to at standard conditions in with and at a of in and to in with of and at for 1 and with of the and to at conditions for 1 with of of and to to and for at standard in for and with for and of stored the in of stored for to months and for a of of RNA in a in one of a of in a and by the for using the for to the using a time for the at storage to the of the of lyophilized and complexed vaccine for the vaccine by a for to of at months for stored with a complexed using the to IntroductionRNA-based vaccines show great promise to effectively address existing and emerging infectious diseases,1Deering R.P. Kommareddy S. Ulmer J.B. Brito L.A. Geall A.J. Nucleic acid vaccines: prospects for non-viral delivery of mRNA vaccines.Expert Opin. Drug Deliv. 2014; 11: 885-899Crossref PubMed Scopus (105) Google Scholar, 2Rauch S. Jasny E. Schmidt K.E. Petsch B. New vaccine technologies to combat outbreak situations.Front Immunol. 2018; 9: 1963Crossref PubMed Scopus (298) Google Scholar, 3Zhang C. Maruggi G. Shan H. Li J. Advances in mRNA vaccines for infectious diseases.Front Immunol. 2019; 10: 594Crossref PubMed Scopus (322) Google Scholar including the ongoing pandemic caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). RNA vaccines can be rapidly adapted to new targets and manufactured using sequence-independent operations, thus reducing the cost and time to develop new vaccines, particularly in pandemic settings.4Pardi N. Hogan M.J. Porter F.W. Weissman D. mRNA vaccines a new era in vaccinology.Nat. Rev. Drug Discov. 2018; 17: 261-279Crossref PubMed Scopus (1516) Google Scholar The Emergency Use Authorization granted to two safe and highly effective mRNA vaccines targeting SARS-CoV-2, less than 1 year after sequencing the novel coronavirus, highlights the power of this new technology.5Jackson L.A. Anderson E.J. Rouphael N.G. Roberts P.C. Makhene M. Coler R.N. McCullough M.P. Chappell J.D. Denison M.R. Stevens L.J. et al.An mRNA vaccine against SARS-CoV-2 - preliminary report.N. Engl. J. Med. 2020; 383: 1920-1931Crossref PubMed Scopus (1675) Google Scholar,6Polack F.P. Thomas S.J. Kitchin N. Absalon J. Gurtman A. Lockhart S. Perez J.L. Perez Marc G. Moreira E.D. Zerbini C. et al.Safety and efficacy of the BNT162b2 mRNA covid-19 vaccine.N. Engl. J. Med. 2020; 383: 2603-2615Crossref PubMed Scopus (6139) Google Scholar However, one of the biggest challenges facing these extraordinary new vaccines is the ability to successfully distribute them widely in the face of a pandemic. Cold chain storage is required for both authorized vaccines (−70°C and −20°C for the SARS-CoV-2 RNA vaccines produced by Pfizer/BioNTech and Moderna, respectively). Frozen shipping and storage even at standard freezer conditions poses difficulties in settings with well-established medical infrastructure, challenges greatly compounded in areas with limited resources.7Kumru O.S. Joshi S.B. Smith D.E. Middaugh C.R. Prusik T. Volkin D.B. Vaccine instability in the cold chain: mechanisms, analysis and formulation strategies.Biologicals. 2014; 42: 237-259Crossref PubMed Scopus (208) Google Scholar, 8Chen D. Zehrung D. Desirable attributes of vaccines for deployment in low-resource settings.J. PubMed Scopus Google Scholar, Volkin D.B. E. the cold of mRNA vaccine PubMed Scopus Google of in RNA vaccines is a the this are and Volkin D.B. E. the cold of mRNA vaccine PubMed Scopus Google Scholar, D. G. nanoparticle vaccines: and J. PubMed Scopus Google Scholar, of storage and of covid-19 9: PubMed Scopus Google Scholar However, are vaccine RNA are to by of the RNA has in to by and a new of Rev. Drug Discov. 2014; PubMed Scopus Google Scholar to and RNA alone a to Anderson in Rev. Drug Discov. PubMed Scopus Google Scholar RNA delivery are to and RNA by et A. Anderson the in technologies for mRNA 2019; PubMed Scopus Google Scholar and et S. J. in delivery of mRNA using delivery PubMed Scopus Google The current system of for RNA vaccines, including SARS-CoV-2 vaccines in to is a lipid nanoparticle delivery L.A. Anderson E.J. Rouphael N.G. Roberts P.C. Makhene M. Coler R.N. McCullough M.P. Chappell J.D. Denison M.R. Stevens L.J. et al.An mRNA vaccine against SARS-CoV-2 - preliminary report.N. Engl. J. Med. 2020; 383: 1920-1931Crossref PubMed Scopus (1675) Google S. Advances in lipid for PubMed Scopus Google Scholar, for 2014; PubMed Scopus Google Scholar, A. D. mRNA vaccine delivery using lipid Deliv. PubMed Scopus Google Scholar, A. Brito L.A. Smith M. J. et and of by mRNA vaccines against and PubMed Scopus Google Scholar in the RNA is a lipid This in that the RNA and by the A. D. mRNA vaccine delivery using lipid Deliv. PubMed Scopus Google K.E. E. A. A. S. J. T. et of lipid for of mRNA Nucleic 2019; PubMed Scopus Google Scholar However, of both the RNA and Volkin D.B. E. the cold of mRNA vaccine PubMed Scopus Google Scholar, D. G. nanoparticle vaccines: and J. PubMed Scopus Google Scholar, of storage and of covid-19 9: PubMed Scopus Google Scholar with to temperatures in to after of lipid the of and J. PubMed Scopus Google J. S. Li G. storage of for mRNA 2020; PubMed Scopus Google Scholar of to the thermostability of RNA vaccines at A. M. D. S. T. T. T. Petsch B. RNA vaccine against 11: PubMed Scopus Google Scholar, Li H. G. C. et mRNA vaccine against 2020; PubMed Scopus Google Scholar, H. Li Li C. G. Li et and efficacy of the mRNA vaccine in PubMed Scopus Google Scholar authorized RNA vaccines in the still Vaccine - - of delivery and to RNA L.A. M. A. T. M. J. A. et for the delivery of RNA 2014; PubMed Scopus Google Scholar, J. B. J. M. E. J. E. S. et nanostructured lipid carrier for delivery of a RNA against 2018; PubMed Scopus Google Scholar, of lipid nanoparticle for and in delivery of 2019; PubMed Scopus Google Scholar the ability of a nanostructured lipid carrier (NLC) system to effectively vaccines by This NLC delivery system can be complexed and lyophilized with The NLC alone for at 1 year of storage at refrigerated while lyophilized NLC/RNA to and ability to protein in after at months of room temperature storage and at months at refrigerated