powerpoint presentationdmsc2.dmsc.moph.go.th/webroot/drug/download... · identification by...
TRANSCRIPT
![Page 1: PowerPoint Presentationdmsc2.dmsc.moph.go.th/webroot/drug/download... · Identification by SDS-PAGE/Western Blotting. SDS-PAGE • Separate by molecular weight. Western Blotting •](https://reader030.vdocuments.mx/reader030/viewer/2022021717/5b542e907f8b9a1f648ca596/html5/thumbnails/1.jpg)
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ANEMIA
http://www.clipartkid.com/red-blood-cell-weak-cliparts/
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http://en.hdbuzz.net/070, http://alportsyndrome.org/blog/stages-of-kidney-disease/, http://www.clipartbest.com/red-blood-cell-cartoon
Erythropoietin
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Recombinant erythropoietin
https://en.wikipedia.org/wiki/Erythropoietin
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Epoetin Erythropoietin
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PRCA
Genetic
Route/Handling
Switching
Quality
Glycosylation
Leachates
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Quality Control for Erythropoietin Products
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Quality
Non-clinical study
Clinical study
Stand-aloneerythropoietin
products
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(BP)
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Dimers and related
substances of higher
molecular weight(Dimer)
Identification(Iden)
pH
BacterialEndotoxin
(Bact)Particulate
Matter(Parti)
Sterility(Ster)
Potency(Poten)
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Albumin
No Albumin
Erythropoietin products in
Thailand
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Ident
• SDS-Page/Western Blotting• Conform to standard
pH• BP• 6.6-7.4
Bact
• BP• Less than 20 IU/ 10000 IU of erythropoietin
Parti
• BP/USP (Harmonization)• ≥ 10 µm not exceed 6000/container and ≥ 25 µm not exceed 600/container
Ster• BP• Sterile
Dimer
• Size Exclusion Chromatography/ SDS-PAGE/Western Blotting• Not greater than 2%
Poten
• Enzyme linked immunosorbent assay (ELISA)• Not less than 80% and not more than 125%
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Identification by SDS-PAGE/Western Blotting
SDS-PAGE• Separate by
molecular weight
Western Blotting
• Transfer to membrane for detection
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SDS-PAGE
https://www.youtube.com/watch?v=IWZN_G_pC8U
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Western Blotting
https://www.youtube.com/watch?v=VgAuZ6dBOfs
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Erythropoietin injection (BP 2016)
ProductionWhere the product contains human serum albumin, itdoes not comply with the test for Dimers and related substances of higher molecular weight; however the manufacturing process is validated to show that aggregation does not occur
Dimers and related substances of higher molecular weight
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Dimers and related substances of higher molecular weight
Aggregate
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Dimers and related substances of higher molecular weight
Size exclusion chromatography
(SEC)
• Prepare 2% aggregate standard
SDS-PAGE/Western blotting
• Semi-quantitate aggregate in sample
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Size exclusion chromatography
(SEC)
High Performance Liquid Chromatography(HPLC)
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Size exclusion chromatography
(SEC)
https://www.youtube.com/watch?v=rPRbqYWlSEo
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Non-reducing
Reducing
SDS-PAGE/Western Blotting
For DIMER
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Potency by ELISA
http://www.cstj.co.jp/ddt/elisa_line.php
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https://www.youtube.com/watch?v=6Ue1Hd3dyaQ
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Quality
Non-clinical study
Clinical study
Stand-aloneerythropoietin
products
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PRCA
Genetic
Route/Handling
Switching
Quality
Glycosylation
Leachates
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Glycosylation for Erythropoietin
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Erythropoietin concentrated
solution (Drug substance)
Erythropoietin product
(Drug product)
Glycosylation
Photos from http://www.process-worldwide.com/single-use-systems-for-flexible-operations-in-biopharmaceutical-production-a-439523/ and http://www.equinechronicle.com/new-withdrawal-period-for-intra-articular-injections-and-shockwave-therapy-in-competition-horses/
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Erythropoietin product
(Drug product)
https://senthilarivan.wordpress.com/2015/12/06/isoelectric-focusing/
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https://www.youtube.com/watch?v=-rDRU_qxwYA and http://aem.asm.org/content/64/7/2572/F1.expansion.html
Erythropoietin concentrated
solution (Drug substance)
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Biosimilar
Drug substance