Simple and Rapid One-Step Elisa for T7 RNA Polymerase Detection

The rapid development of mRNA-based vaccines and therapeutics has opened up groundbreaking possibilities in modern medicine. However, ensuring the safety, efficacy, and regulatory compliance of mRNA drugs requires precise quality control at every stage of development. One critical parameter is the detection of residual T7 RNA polymerase, an enzyme commonly used during in vitro transcription of mRNA.

To address this need, Hzymes has developed the T7 RNA Polymerase ELISA Kit — a simple, one-step, highly specific, and sensitive assay designed to meet international quality guidelines, including the FDA’s draft guidance on mRNA vaccine quality analysis (2023).

Application

This kit detects the residual amount of T7 RNA polymerase by double antibody sandwich enzyme-linked immunosorbent assay (ELlSA) in accordance with the recommended method in the "mRNA Vaccine Quality Analysis Method- Draft Guidance".

Table2. characterization and release testing for mRNA Drug Substance

Advantages

1.     Easy to operate

One step protocol with sample-to-result in 75min.

2.     Excellent Adaptability

Standard Curve: R 2> 0.99

Detection Limit <0.1 ng/ml

Limit of Quantification: 0.25 ng/ml

High accuracy and Repeatability: CV <10% with spike recovery in the range of 80%-120%.

3. High Specificity

No cross-reactivity to other proteins.

4. Good adaptation

Adapt to different T7 RNA polymerase from multiple suppliers.

Performances

1 Limit of Detection (LOD) as low as 0.1ng/ml and Limit of Quantification (LOQ) as low as 0.25 ng/ml:

2 High accuracy and Repeatability: CV <10% with spike recovery in the range of 80%-120%.

3 High Specificity: no cross-reactivity to other proteins

The T7 RNA polymerase Elisa kit only specifically recognizes the T7 RNA polymerase with no cross-reactivity to other lVT tool enzymes.

4 Good adaptation:

Current T7 RNA Polymerase ELISA kit from suppler A, B, C, D doesn't match with multiple T7 RNA polymerase on the market.

Hzvmes' T7 RNA Polymerase ELlSA kit is adapt to different T7 RNA polymerase from multiple suppliers.

5 mRNA Sample Measurement

T7 RNA polymerase residues detection was performed for actual mRNA samples with the concentration 1-2mg/mlobtained from different CMC process stages.

Note: Sample B was spiked at the maximum dilution factor of 4ng/ml, 2ng/ml and 1ng/ml, respectively, Samples A and Care in each the dilution factor was spiked at 4ng/ml.

6 Good Reproducibility: Mouse Epo mRNA sample at 1 mg/ ml concentration was measured for 3 times.

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