Streamlined and on-demand preparation of mRNA products on a universal integrated platform

Vaccines are used to protect human beings from various diseases.mRNA vaccines simplify the development process and reduce the production cost of conventional vaccines,making it possible to respond rapidly to acute and severe diseases,such as coronavirus disease 2019.In this study,a universal integrated platform for the streamlined and ondemand preparation of mRNA products directly from DNA templates was established.Target DNA templates were amplified in vitro by a polymerase chain reaction module and transcribed into mRNA sequences,which were magnetically purified and encapsulated in lipid nanoparticles.As an initial example,enhanced green fluorescent protein(eGFP)was used to test the platform.The expression capacity and efficiency of the products were evaluated by transfecting them into HEK-293T cells.The batch production rate was estimated to be 200–300μg of eGFP mRNA in 8 h.Furthermore,an mRNA vaccine encoding the receptor-binding domain(RBD)of the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)spike protein was produced by this platform.The proposed integrated platform shows advantages for the universal and on-demand preparation of mRNA products,offering the potential to facilitate broad access to mRNA technology and enable the development of mRNA products,including the rapid supply of new mRNA-based vaccines in pandemic situations and personalized mRNA-based therapies for oncology and chronic infectious diseases,such as viral hepatitis and acquired immune deficiency syndrome.

Monkeypox virus quadrivalent mRNA vaccine induces immune response and protects against vaccinia virus

Monkeypox has been declared a public health emergency by the World Health Organization.There is an urgent need for efficient and safe vaccines against the monkeypox virus(MPXV)in response to the rapidly spreading monkeypox epidemic.In the age of COVID-19,mRNA vaccines have been highly successful and emerged as platforms enabling rapid development and large-scale preparation.Here,we develop two MPXV quadrivalent mRNA vaccines,named mRNA-A-LNP and mRNA-B-LNP,based on two intracellular mature virus specific proteins(A29L and M1R)and two extracellular enveloped virus specific proteins(A35R and B6R).By administering mRNA-A-LNP and mRNA-B-LNP intramuscularly twice,mice induce MPXV specific IgG antibodies and potent vaccinia virus(VACV)specific neutralizing antibodies.Further,it elicits efficient MPXV specific Th-1 biased cellular immunity,as well as durable effector memory T and germinal center B cell responses in mice.In addition,two doses of mRNA-A-LNP and mRNA-B-LNP are protective against the VACV challenge in mice.And,the passive transfer of sera from mRNA-A-LNP and mRNA-B-LNP-immunized mice protects nude mice against the VACV challenge.Overall,our results demonstrate that mRNA-A-LNP and mRNA-B-LNP appear to be safe and effective vaccine candidates against monkeypox epidemics,as well as against outbreaks caused by other orthopoxviruses,including the smallpox virus.