Jag tar det igen, regulatoriska lagar kontrollerar tester och grnsvrden. Om ngon produkt ( i det hr fallet batch) inte klarar kraven slpps inte batchen. Det avsnitt DU citerar (2.2.6. Recommendations for future quality development) i ditt tidigare inlgg avser inte produkten utan de material och komponenter som anvnds i produktionen dr man freslr frbttringar. Det gr man i princip alltid vid alla typer av inspektioner och prvningar. Det pverkar inte ngra grnsvrden. Batcher och produkter testas alltid flera gnger av producent, regulatorisk myndighet och mottagare. Om ngot resultat skiljer sig frn tidigare eller om ett test inte klarar kravet slpps inte batchen/produkten.
Nej, det har inte funnits ngra problem med ngon batch som slppts. Du fr LSA det dokument du refererar till s framgr exakt vad som gller.
Ja, det str i dokumentet vad som sklijer Process 1 och Process 2 - du fr LSA dokumentet s framgr det. LS sid 18-19. Pajas.
Dina tidigare kommentarer om de "variationer" som man noterat betyder att man vill veta varfr olika processer producerar olika resultat. Den variationen pverkar inte godknnandet d vrdena ligger inom tilltna niver och produkten ligger inom alla grnsvrden. Det str ocks i det dokument du INTE lnkade till. Jag fr hjlpa dig:
https://www.ema.europa.eu/en/documen...-report_en.pdf
De tester som grs och den metod som anvnds skiljer sig inte mellan FDA, EMA eller ngon annan regulatorisk myndighet. Det r samma ver hela linjen (beroende p att i princip alla regulatoriska myndigheter har kopierat FDAs). Jag skall vara snll mot dig och lista de tester och metoder som anvnds. Testerna delas upp i tre avsnitt:
1. Test av Plasmid DNA
2. mRNA Drug Substance testing (batch)
3. Release test of product (man testar fyllda vials)
Hr r de tester som utfrs fr att verifiera plasmid DNA:t:
1A. Identity: mRNA sequence identity confirmation. Metod: Sanger sequencing + RT-PCR
1B. Identity: Lipid Identity. Metod: RP-HPLC-CAD
1C. Content: RNA concentration/RNA encapsulation efficiency. Metod: Fluorescence-based assay
1D. Content: Lipid Content. Metod: RP-HPLC-CAD
1E. Integrity: LNP size and polydispersity. Metod: DLS
1F. Integrity: RNA size and integrity. Metod: CGE
1G. Purity: Product related impurities - aggregate quantitation. Metod: SEC-HPLC
1H. Purity: Product related impurities - percentage of fragment mRNA. Metod: IP-RP-HPLC
1I. Potency: Expression of target protein. Metod: Cell-based essay
1J. Safety: Endotoxin. Metod: Gel Clot + Turbidimetric + Chromogenic
1K. Safety: Bioburdin. Metod:Peptid
1L. Safety: Sterility. Metod:Nucleic acid amplification + direct transfer
1M. Generic: Appearance. Metod: Light Obscuration Particle Count
1N. Generic: Ph.
Hr r de tester som utfrs fr att verifiera batcher:
2A. Identity: mRNA sequence identity confirmation. Metod: HTS
2A. Identity: mRNA sequence identity confirmation. Metod: Sanger sequencing
2A. Identity: mRNA sequence identity confirmation. Metod: RT-PCR
2B. Content: RNA concentration. Metod: qPCR
2B. Content: RNA concentration. Metod: dPCR
2B. Content: RNA concentration. Metod: Ultraviolet Spectroscopy
2C. Integrity: mRNA intactness. Metod: Capillary electrophoresis
2C. Integrity: mRNA intactness. Metod: Capillary gel electrophoresis (CGE)D
2C. Integrity: mRNA intactness. Metod: Agarose gel electrophoresis
2D. Purity: 5 capping efficiency. Metod: Reverse-phase liquid chromatography mass spectroscopy (RPLC-MS/MS)
2D. Purity: 5 capping efficiency. Metod: Ion pair reversed-phase high-performance liquid chromatography (IP-RP-HPLC)
2D. Purity: 3 poly(A) tail length. Metod: Ion pair reversed-phase high-performance liquid chromatography (IP-RP-HPLC)
2D. Purity: Product related impurities - dsRNA. Metod: Immunoblot
2D. Purity: Product related impurities - dsRNA. Metod: Enzyme-linked immunosorbent assay (ELISA)
2D. Purity: Product related impurities - aggregate quantitation. Metod: Size exclusion-high-performance liquid chromatography (SEC-HPLC)
2D. Purity: Product related impurities - percentage of fragment mRNA. Metod: Reversed-phase HPLC (RP-HPLC)D
2D. Purity: Process related impurities-residual DNA template. Metod: qPCR
2D. Purity: Process related impurities - quantitation of free/ non-incorporated nucleosides. Metod: Reverse-phase liquid chromatography mass spectroscopy (RPLC-MS/MS)D
2D. Purity: Process related impurities - residual T7 RNA polymerase content. Metod: Enzyme-linked immunosorbent assay (ELISA)
2E. Potency: Expression of target protein. Metod: Cell-based essay
2F. Safety: Endotoxin. Metod: Gel Clot + Turbidimetric + Chromogenic
2F. Safety: Bioburdin. Metod: Peptid
2G. Generic: Appearance. Method: Light Obscuration Particle Count
2G. Generic: Residual Solvents. Method: Gas Chromatography
2G. Generic: pH.
Hr r de tester som utfrs fr att verifiera produkter (vials):
3A. Identity: mRNA sequence identity confirmation. Metod: Sanger sequencing
3A. Identity: mRNA sequence identity confirmation. Metod: RT-PCR
3A. Identity: Identity of lipids. Metod: Reversed-phase high-performance liquid chromatography with charged aerosol detector (RP-HPLC-CAD)
3B. Content: RNA concentration. Metod: Fluorescence-based assay
3B. Content: RNA encapsulation efficiency. Metod: Fluorescence-based assay
3B. Content: Lipid Content. Metod: Reversed-phase high-performance liquid chromatography with charged aerosol detector (RP-HPLC-CAD)
3C. Integrity: LNP size and polydispersity. Metod: Dynamic light scattering (DLS)
3C. Integrity: RNA size and integrity. Metod: Capillary gel electrophoresis (CGE)
3D. Purity: Product related impurities - aggregate quantitation. Metod: Size exclusion-high-performance liquid chromatography (SEC-HPLC)
3D. Purity: Product related impurities - percentage of fragment mRNA. Metod: Ion pair reversed-phase high-performance liquid chromatography (IP-RP-HPLC)
3E. Potency: Expression of target protein. Metod: Cell-based essay
3F. Safety: Endotoxin. Metod: Gel Clot + Turbidimetric + Chromogenic
3F. Safety: Sterility. Metod: Nucleic acid amplification + direct transfer
3G. Generic: Appearance. Method: Light Obscuration Particle Count
3G. Generic: Residual Solvents. Method: Gas Chromatography
3G. Generic: Osmolality. Method: FPDT
3G. Generic: Subvisible particles. Method:LIGHT OBSCURATION PARTICLE
3G. Generic: Extractable volume.
3G. Generic: Container Closure Integrity.
Vill du veta hur componenter, batcher och produkter testas samt vilka lagar som styr freslr jag:
https://www.ecfr.gov/current/title-2...ter-F/part-600
Vill du veta vilka tester som genomfrs freslr:
https://www.usp.org/
Vill du veta mer om de standarder och regler som styr testningen tills dess att den lmnar producent freslr jag:
http://ftp.uspbpep.com/
Dessa tester utfrs av bde producent, regulatorisk myndighet samt tredje part. Om ngon av testerna visar annorlunda resultat stoppas batchen. Produkten testas igen av den regulatoriska myndigheten i mottagarlandet samt tredjepart. terigen, om ngot resultat skiljer sig frn andra resultat i batchens eller produktens historia s stoppas batchen.
Vill du ha resultaten frn de tester som genomfrs fr du kontakta EMA enligt fljande guide och gra en begran:
https://www.ema.europa.eu/en/documen...cuments_en.pdf