Removing the immunogenic dsRNA that standard mRNA purification leaves behind

Removing double-stranded RNA from mRNA drug substance is a manufacturing problem with no easy answers. dsRNA is an unavoidable byproduct of in vitro transcription, fully triggers innate immune sensing at trace levels, and overlaps the mRNA product in size, charge, and poly(A) content, so conventional purification lets it through.
Here is how Repligen built a dsRNA-selective affinity capture step, engineering the ligand and bead, screening binding conditions in a 96-well filter plate, and validating on a column, cutting dsRNA by >100-fold at ~83% mRNA recovery and producing material, made with standard unmodified nucleotides.

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