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. 2022 Aug 4;29:618–624. doi: 10.1016/j.omtn.2022.08.001

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This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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Evaluating mRNA integrity, dsRNA content, protein expression, and immuno-stimulatory effect for mRNA1, mRNA2, and HPLC mRNA

(A) Agarose gel with RNA ladder. (B) J2 mAb dot blot. (C) Protein expression, the mRNA dose is 200 ng, and protein expression from mRNA2 was defined as 100% in this example. (D) immuno-stimulatory (IS) response; the mRNA dose is 1,250 ng; IS response triggered from HPLC mRNA was defined as 1 in this example. All experiments were repeated three times, and the error bars are standard deviation of the mean. mRNA1 is purified mRNA produced from regular IVT, mRNA2 is purified mRNA produced from IVT with 1.2 M urea, and HPLC mRNA is the mRNA produced from regular IVT and purified by RP-HPLC. Polyinosinic:polycytidylic acid [Poly(I:C)], a synthetic dsRNA was used as positive control in the assay.