Development and scale-up of oligo-dT monolithic chromatographic column for mRNA capture through understanding of base-pairing interactions

•Oligo-dA binding assay was developed for characterization of monolith OdT columns.•Coupling ≥ 2.5 probes of 18-mer OdT per 100 nm2 is optimal for mRNA chromatography.•Increased mRNA residence time is needed to utilize the full capacity of OdT column.•Up to 5 mg/mL mRNA dynamic binding capacity was achieved at 1 min residence time.•Oligo-dT monolith column enables efficient gram-scale mRNA purification.