Cell-based Potency Assays
The Significance of mRNA Potency Assay
With the rapid development of mRNA vaccines and therapeutics, a specific panel of analysis methods is needed to test the potency (defining their biological activity), involving the translational efficiency of mRNA. These methods are typically based on cell transfection and functional assays of the target protein.
Yaohai Bio-Pharma offers cell-based potency assays for mRNA drug substances or LNP products, including cell transfection (293T, HepG2, HeLa, K562, U937, Raw264.7 cells, etc.) and ELISA or Western Blotting (WB) assays.
Regulatory Requirements for Potency Assays
According to the WHO regulatory considerations, “The potency of each lot of the final vaccine should be determined using a suitably quantitative and validated functional method(s). Potential in vitro potency assays may include cell-based transfection systems or cell-free assays.” And the USP recommended a “cell-based assay” to test the potency of mRNA, i.e., expression of the target protein.
Analytical Method
|
Analysis
|
Methods
|
|
Potency Assays
|
Cell-based transfection and target protein identify:
293T, HepG2, HeLa, K562, U937, Raw264.7 cells, etc.
|
Procedure
Step 1. Cell-based Transfection with mRNA or mRNA-LNP
The cells are cultured and transfected with mRNA or mRNA-LNP. In detail, mRNA is transfected after being mixed with a transfection reagent, while mRNA-LNP is directly transfected into the cells. After 24-48 hours, clarify the supernatant containing the expressed protein by centrifugation.
Step 2. Functional Assays of the Target Protein
Perform ELISA or WB assays to assess the functional binding of the target proteins.
Case Study
Cell-based Potency Assays of mRNA Samples
Yaohai-BioPharma has developed cell-based potency assay method for mRNA, involving cell culture, cell transfection and protein functional assays. We transfected eGFP mRNA, mCherry mRNA, luciferase mRNA, spike protein mRNA into 293T cells, and detected target protein through fluorescence, luminescent or colored signal, WB, ELISA.
Cell-based Potency Assays of mRNA Products by Yaohai Bio-Pharma
EN
AR
HR
CS
DA
NL
FI
FR
DE
EL
IT
JA
KO
NO
PL
PT
RO
RU
ES
SV
IW
ID
LV
LT
SR
SK
SL
UK
VI
ET
HU
TH
TR
FA
AF
MS
BE
MK
UR
BN
