Journal: Organic Process Research & Development

Article Title: Continuous Purification of a Conjugated Short Interfering RNA Therapeutic Using Anion Exchange Twin-Column Chromatography (MCSGP)

doi: 10.1021/acs.oprd.4c00513

Figure Lengend Snippet: Figure 1. Twin column chromatography system setup for MCSGP (Contichrom CUBE, YMC ChromaCon).

Article Snippet: Tables S2 and S3 detail specific MCSGP method parameters used to run “startup” and “main” methods on the Contichrom CUBE hardware.

Techniques: Column Chromatography

Journal: Organic Process Research & Development

Article Title: Continuous Purification of a Conjugated Short Interfering RNA Therapeutic Using Anion Exchange Twin-Column Chromatography (MCSGP)

doi: 10.1021/acs.oprd.4c00513

Figure Lengend Snippet: Figure 2. MCSGP process. (A) An MCSGP run with 4 cycles is shown as an example. A short startup step is needed to preload column 1 with feed before the main cyclic phase of MCSGP begins. Each cycle consists of an elution from column 1 (switch 1red) and an elution from column 2 (switch 2blue), respectively. Alternating elutions can continue indefinitely for “n” cycles until the feed is consumed. To finish the MCSGP run, a shutdown step consisting of a single elution without any refeeding is done. (B) A detailed schematic representation of a single MCSGP “switch” is shown. This consists of four phases: P1 = phase 1 (elution + regeneration); P2 = phase 2 (elution with weak recycling); P3 = phase 3 (product collection + refeeding); and P4 = phase 4 (elution with strong recycling). Columns are operated in parallel during P1 and P3; columns are interconnected for product recycling in P2 and P4. The position of the columns alternates switch to switch, allowing for continuous operation (yellow arrows indicate column interchange between switches). See Section 2.4 for a more complete description of the MCSGP process principle.

Article Snippet: Tables S2 and S3 detail specific MCSGP method parameters used to run “startup” and “main” methods on the Contichrom CUBE hardware.

Techniques:

Journal: Organic Process Research & Development

Article Title: Continuous Purification of a Conjugated Short Interfering RNA Therapeutic Using Anion Exchange Twin-Column Chromatography (MCSGP)

doi: 10.1021/acs.oprd.4c00513

Figure Lengend Snippet: Figure 3. Single-column AIEX chromatographic profiles and the MCSGP design. (A) Black line: representative chromatogram of pilot-scale single- column batch production run showing absorbance UV@290 nm vs column volumes (CV) overlaid with %B modifier concentration (purification using the AKTA pure 150 M). The pilot-scale method was downscaled 78.6× and run on the Contichrom CUBE. (B) MCSGP design chromatogram. Absorbance UV@280 nm is overlaid with product purity (area% HPLC-UV). Product purity was measured by HPLC analysis, and each dot represents an analyzed fraction. Phases P1 to P4, as described in Figure 2B, are highlighted showing where MCSGP recycling and product collection boundaries were set. (C) - The MCSGP Wizard v8.1 software simplifies method creation.

Article Snippet: Tables S2 and S3 detail specific MCSGP method parameters used to run “startup” and “main” methods on the Contichrom CUBE hardware.

Techniques: Concentration Assay, Purification, Software

Journal: Organic Process Research & Development

Article Title: Continuous Purification of a Conjugated Short Interfering RNA Therapeutic Using Anion Exchange Twin-Column Chromatography (MCSGP)

doi: 10.1021/acs.oprd.4c00513

Figure Lengend Snippet: Figure 4. (A, B) MCSGP chromatographic profiles showing absorbance UV@280 nm and %B (A) or UV@280 nm and conductivity (mS/cm) (B) plotted vs time (min). (A) Plot showing 13 consecutive MCSGP cycles including startup and shutdown (total of 27 product elutions). (B) Plot showing an overlay of 13 consecutive MCSGP cycles. (C, D) Plots showing the total area under the curve (AUC) in (mAU×mL) vs cycle number. Each dot is the sum of 2 elutions from 2 switches. (C) Black line: total AUC. Red line: AUC for the product collection window. (D) Blue line: AUC for the weak recycling phase. Green line: AUC for the strong recycling phase.

Article Snippet: Tables S2 and S3 detail specific MCSGP method parameters used to run “startup” and “main” methods on the Contichrom CUBE hardware.

Techniques:

Journal: Organic Process Research & Development

Article Title: Continuous Purification of a Conjugated Short Interfering RNA Therapeutic Using Anion Exchange Twin-Column Chromatography (MCSGP)

doi: 10.1021/acs.oprd.4c00513

Figure Lengend Snippet: Figure 5. MCSGP product quality evaluation. (A) Plot showing cycle to cycle MCSGP target product yield (%) and mass recovered (mg). (B) Plot showing cycle to cycle product purity HPLC-UV (area%) vs impurity N-1 content HPLC-UV (area%). (C) Analytical HPLC chromatograms (UV Abs at 260 nm, normalized and then zoomed in to show impurities). Blue line is the crude feed material (85 area% purity). Orange line is the 13 cycle MCSGP product pool (95.4 area% purity).

Article Snippet: Tables S2 and S3 detail specific MCSGP method parameters used to run “startup” and “main” methods on the Contichrom CUBE hardware.

Techniques: