The capping rate is an important quality attribute indicator of mRNA vaccine drugs and a necessary validation document for the application of mRNA products. At present, the determination of capping rate is mainly carried out by liquid chromatography-mass spectrometry. There is a difference in molecular weight between capped mRNA and uncapped mRNA in the order of hundreds. Compared with the molecular weight of intact mRNA in the hundreds of thousands, it is difficult to precisely analyze the difference between capped and uncapped samples. Therefore, the analysis of the capping rate first requires processing long mRNA fragments into small ones, transforming the differences of the hundreds of thousands into those of the hundreds of thousands.
Enzymatic capping ↓
Co-transcription capping↓
The mRNA capping rate detection pretreatment kit developed by Baorui Biotechnology can precisely and efficiently process mRNA samples. The processed samples can be directly used for liquid chromatography-mass spectrometry analysis, facilitating the rapid analysis of sample capping rates. This kit is suitable for the pretreatment of mRNA samples with different capping methods and various cap structures.
1Kit components
Reagent Name Volume
Adsorption magnetic beads250μL
Mreagent12μL
10×M buffer50μL
Loading buffer50μL
Buffer 1 1mL
Buffer 2 10mL
Buffer 3 1mL
EB buffer 1mL
★ It can be used for the processing of mRNA produced by different capping processes
★ It can be used for processing mrnas with different types of cap structures
It can be used for sample processing in different models of liquid chromatography-mass spectrometry systems
Convenient, fast, efficient and time-saving
4Probe design and synthesis
SDS-PAGE detection was performed after enzymatic digestion. The system after enzymatic digestion contains probe bands, target fragment bands and other fragments. Among them, the target fragment has a clear band, and its content is basically consistent with that of the probe, significantly higher than that of other fragments, indicating that the enzymatic digestion process was properly handled, and the sample can be used for the next recovery operation.
The fragments after impurity washing and recovery were detected by SDS-PAGE. All fragments except the target band and the probe band had been washed clean. The presence of the probe in the sample will not affect the subsequent liquid chromatography-mass spectrometry (LC-MS) determination. On the contrary, the probe band with a known molecular weight can be used as a scale for subsequent determination.
7Liquid chromatography-mass spectrometry determination and analysis
liquidChromatogram
The total ionic liquid of the sample peakSpectral graph
Cap0 and unCap ion flow mass spectrometry analysis
Liquid chromatography-mass spectrometry determination and analysis
Liquid chromatogram
Total ion flow mass spectrometry of the sample peak
Ion flow mass spectrometry analysis of Cap1 and Cap0
Attached: Product Catalogue
