mRNAs transcribed in vitro by T7 RNA polymerase may contain various contaminants, such as short RNAs produced by abortive initiation events, double-stranded (ds)RNAs generated by self-complementary 3’extension, as well as unincorporated nucleoside triphosphates, small abortive transcripts and plasmid template. Certain RNA sequences even induce high levels immunogenicity.
APExBIO offers purification service to remove the contaminants of modified nucleotide-containing mRNA, thus increase the processing efficiency for downstream applications.
Silica-gel Membrane Spin Column Purification:
It is a solid phase extraction technique for fast nucleic acid purification. mRNA can be bound to solid phase of silica-gel membranes under certain conditions, with subsequent washing and elution steps in water or TE pH 7. This method eliminates most proteins, DNA and NTPs.
HPLC purification by ?KTA avant system:
mRNA can be purified by HPLC (?KTA avant system) using column matrix of alkylated non-porous polystyrene-divinylbenzene copolymer microspheres and optimized buffer system, followed by mRNA analyses and mRNA isolation from column fractions.
HPLC purification removes dsRNA and other contaminants from in vitro synthesized modified nucleotide-containing mRNAs, yielding mRNA with the high level of translation without generation of immunogenicity or RNA sensor activation.
mRNA and long RNA products
APExBIO supplies the best quality mRNA and long RNA. This new product lines involve custom synthesis of mRNA and long RNA (up to multiple kilobases) with a wide array of modification services at scales ranging from micrograms to milligrams. The mRNA can be generated from DNA templates provided by our customers or we can provide a full service from the ground up. We offer mCAP or ARCA capping or modified nucleotides implication for all our standard mRNA transcripts.
All of our mRNA products offer:
Incorporates an anti-reverse cap analog (ARCA) into the transcript to increase translation efficiency
Reduces host cell immune response and enhances stability by incorporating modified nucleotides (5mCTP and ψUTP) and a poly(A) tail
Degrades the DNA template after RNA synthesis with DNase
Removes the 5’ triphosphates at the end of the RNA with phosphatase to further reduce innate immune responses in mammalian cells
Employs a robust clean-up spin column system that delivers high yields of mRNAs that are ready for most downstream applications