Chii chinonzi mRNA vaccine
Iyo mRNA yekudzivirira inoendesa RNA kumaseru emuviri kuratidza uye kugadzira maprotein antigens mushure mekugadziriswa kwakakodzera mu vitro, nekudaro zvichitungamira muviri kuburitsa mhinduro yekuzvidzivirira kubva kune antigen, nekudaro kuwedzera simba remuviri rekuzvidzivirira.[1,3].
Mufananidzo 1: Schematic diagiramu yemhedzisiro yejekiseni yakananga yekudzivirira mRNA [2]
Kurongwa kwemRNA vaccines
MRNA vaccine yakakamurwa kuita marudzi maviri:isingaenzaniswimRNA uyekuzvisimudziramRNA: self-amplifying mRNA haingoverengeri inotarirwa antigen, asiwo inokodha iyo kudzokorora iyo inogonesa intracellular RNA kukwidziridzwa uye mapuroteni ekutaura maitiro.Mishonga isingadzokeri yemRNA inongovharira maantigen ekunangwa uye ine 5'and 3'untranslated regions (UTR).Vanopa kurudziro yakazara yekuchinjika uye kusadzivirirwa kwemukati, zvinoti in situ antigen kutaura uye kutapurirana kwengozi chiratidzo, uye vane zvinotevera maapplication[2,3]
● Inogona kupa kurudziro yakakwana yekuchinjika uye kusadzivirirwa kwemukati, zvinoti in situ antigen kutaura uye kutapurirana kwechiratidzo chengozi.
● Inogona kuunza "yakadzikama" mhinduro yekudzivirira kwemuviri, kusanganisira humoral uye cellular madhara uye immune memory
● Inogona kusanganisa maantigen akasiyana pasina kuwedzera kuoma kwekugadzirwa kwejekiseni
● Kuenderera mberi kwekuvandudzwa kwesimba rekudzivirira muviri kunogona kuwanikwa kuburikidza nekudzokororwa kwejekiseni, uye hapana kana zvishoma mhinduro yekudzivirira kune mutakuri.
● Mishonga yekudzivirira yemRNA yakagadzikana inogona kurerutsa kutakurwa nekuchengetedza majekiseni
Mufananidzo 2: Schematic diagram ye mRNA vaccine uye yayo antigen kutaura maitiro [4]
Zvimiro zveMRNA vaccines
Kuenzaniswa nemajekiseni echinyakare, mRNA vaccine ine nzira dzakareruka dzekugadzira, kukurumidza kukurumidza kukurumidza, hapana kudiwa kwesero tsika, uye mutengo wakaderera.Kuenzaniswa neDNA vaccines, mRNA vaccines haidi kupinda munyucleus uye hapana njodzi yekubatanidzwa mune iyo host genome.Iyo hafu yehupenyu inogona kugadziriswa nekugadziriswa.
Tafura 1: Zvakanakira uye kuipa kwemRNA vaccines
|
| Advantage | Kuperevedza |
| mRNA vaccine | Kukurumidza kutsvaga nekusimudzira, kugadzirwa kwejekiseni kunongotora mazuva makumi mana | Tamba mhinduro isina kufanira immune system
|
| mRNA kusagadzikana pasi pemamiriro emuviri, zviri nyore kudzikisira | Hazvizosanganisire mujenome kudzivirira zvinogona kurapa shanduko
| |
| Hapana chikonzero chechiratidzo chenyukireya chenzvimbo, chinyorwa | Kushanda kwekuchengetedza nyukireya kunoramba kuchiongororwa
|
Mufananidzo 3: Kuyerera chati kwemRNA kugadzirwa kwekudzivirira uye kugadzirira [4]
Foregene Viral RNA Isolation kit
RT-qPCR Nyore (Imwe Danho)
Nzira dzakavandudzwa dzekugadzirira kwemRNA vaccines
Nekuda kwekusagadzikana kwakashata kwemRNA pachayo, kuderedzwa kuri nyore nemanucleases mumatissue, kuderera kwesero yekupinda, uye kushomeka kweshanduro, kukanganisa uku kunodzikamisa kushandiswa kweMRNA vaccines.Kushandura zvakanaka kunoitawo basa rakakosha.Mota dzekutakura dzinogona kukamurwa kuita mavhairasi mavhairasi uye asiri mavhairasi mavhairasi (kusanganisira liposomes, asiri-liposomes, mavhairasi, nanoparticles, nezvimwewo).Nokudaro, matanho ekuvandudza akakodzera anodiwa.Inotevera inzira yekuvandudza yemishonga yekugadzirira mRNA[2]
1 Synthesize cap analogs kana kushandisa capping enzymes kudzikamisa mRNA uye kuwedzera mapuroteni kududzira nekusunga kune eukaryotic kududzira initiation factor 4E (EIF4E)
2 Gadzirisa zvinhu zviri munharaunda ye5'-isina kushandurwa (UTR) uye 3'-UTR kudzikamisa mRNA uye kuwedzera mapuroteni kududzira.
3 Kuwedzera Poly(A) muswe unogona kudzikamisa mRNA uye kuwedzera mapuroteni kududzira
4 Modified nucleosides kuderedza innate immune activation uye kuwedzera dudziro
5 Kurapa neRNase III uye nekukurumidza protein liquid chromatography (FPLC) kuchenesa kunogona kuderedza immune activation uye kuwedzera kududzira.
6 Natsiridza nhevedzano kana makodoni kuti uwedzere shanduro
7 Kuendesa pamwe chete kwekutanga kweshanduro uye dzimwe nzira dzekushandura shanduro uye immunogenicity
Mufananidzo 4: In vitro transcription (IVT) mRNA kugadzirwa uye gungano maitiro [5]
Kugadzirira kwakakura kweplasmid DNA
Kucheneswa kwePlasmid DNA kunonyanya kubvisa zvinosvibisa zvakaita seRNA, open-denderedzwa DNA endotoxin, mapuroteni anotambira uye ane nucleic acid, uye kazhinji anoshandura recombinant plasmid kuita E. coli.E. coli inopinda mukuvirisa kwepamusoro-density, ipapo kuparadzaniswa kwakasimba-mvura, uye kuunganidzwa kweE. coli.E. coli inobva yaiswa kune alkaline lysis, centrifugal solid-liquid separation uye microfiltration clarification mushure me lysis, ultrafiltration uye concentration mushure mekujekesa, uye ipapo chromatographic purification.
Kucheneswa kweplasmid DNA:
Foregene General Plasmid Mini Kit
【1】苗鹤凡, 郭勇, 江新香.mRNA疫苗研究进展及挑战[J].免疫学杂志, 2016(05):446-449.
【2】Pardi N , Hogan MJ , Porter FW , et al.mRNA majekiseni - nguva itsva mujekiseni[J].Masikirwo Ongororo Kuwanikwa Kwezvinodhaka, 2018.
【3】Kramps T., Elbers K. (2017) Nhanganyaya yeRNA Vaccines.Mu: Kramps T., Elbers K. (eds) RNA Vaccines.Methods in Molecular Biology, vol 1499. Humana Press, New York, NY.
【4】Maruggi G , Zhang C , Li J , et al.mRNA seShanduro yeTekinoroji yeBaccine Development yekudzora Zvirwere Zvinotapukira[J].Molecular Therapy, 2019.
【5】Sergio Linares-Fernández, Céline Lacroix, ,Tailoring mRNA Vaccine to Balance Innate/Adaptive Immune Response,Trends in Molecular Medicine,Volume 26, Issue 3,2020,Mapeji 311-323.
Nguva yekutumira: Aug-05-2021
