Chromacon

MCSGP for Oligonucleotide Purification

Production of oligonucleotides

The production of oligonucleotides for therapeutic use requires that any undesired byproducts of synthesis be removed in order to obtain pharmaceutical grade purity. To this end, a chromatographic polishing step is routinely used, but this typically reduces the overall yield of production by 40-60%. Given the high cost of synthesis, poor chromatographic performance downstream has a major negative impact on the economics of the overall production. As countermeasure, lower purity “waste” products can undergo re-chromatography, however this greatly reduces process productivity and significantly increases the burden of analytical characterization of product fractions. Thus, manufacturers are looking for alternative, more advanced chromatographic methodology to boost yield.

Application Note (AIEX)

​Publication (Reversed Phase)

Economic benefits of the continuous chromatography process MCSGP

The continuous chromatography process MCSGP leads to higher yields at target purity.

MCSGP is a chromatographic method operated continuously using two columns, compared to the batch mode of single-column chromatography (see above). Much of the basic principle, however, remains the same. In a chromatographic process, the impure feed mixture is separated, giving fractions with pure product, fractions with a mixture of impurities and product, and fractions containing only impurities. Pure product fractions are collected and impure product-containing side fractions are discarded or re-processed. If product containing side fractions are discarded, valuable product is lost. Reprocessing the side fractions accumulates impurities and applicability is thus limited. Moreover, reprocessing reduces overall productivity. MCSGPcan isolate most of the product included in the starting material by internally recycling product-containing side fractions and repeatedly separating impurities and pure product. The recycling is done automatically, without the need for off-line analysis. The impure side fractions are transferred from one chromatographic column to another. Fresh feed material is added every cycle. This ensures continuous operation and leads to a product with high yield without compromising purity.

Purification of MCSGP with Contichrom technology

MCSGP offers significant advantages over single-column chromatography in the purification of oligonucleotides. The advantages include:

  • Oligonucleotide yields were increased by at least 50%, leading to over 90% recovery at 92% purity (target purity was >91%)
  • For single-column chromatography, the recovery was 60% at 92% purity
  • Increased yield allows massive scale-down of oligonucleotide synthesis upstream to obtain required oligo amounts
  • The productivity was increased 2-fold compared to the Benchmark run
  • A massive reduction in the number of samples for analytical characterization, especially if re-chromatography of waste fractions is carried out. (Only one sample is generated per MCSGP cycle for two feed injections, while multiple fractions per injection are generated in single-column chromatography)
  • The high yield of MCSGP makes re-chromatography obsolete
  • All other process parameters were comparable to the batch runs

The increase in productivity by MCSGP allows multiple manufacturing options:

  • The same target amount can be produced within the same time with smaller columns
  • A specified target amount can be produced in a shorter period of time with columns of the same size
  • More target compound can be produced per total column volume in the same time

Contichrom CUBE 

The MCSGP process can be operated by all Contichrom ® CUBE  systems. The Contichrom ® CUBE  is a versatile preparative laboratory-scale chromatography system for single- and twin-column processes with 100 bar (1450 psi) pressure rating. ChromIQ ® , the operating software of Contichrom ® systems, contains a wizard for designing and operating the MCSGP process.


 

Contichrom ® TWIN HPLC GMP production scale-up systems

With the Contichrom ® TWIN HPLC series from YMC, MCSGP is available for manufacturing under GMP conditions. The twin-column scale-up systems have been co-developed by YPT and ChromaCon to ensure transferability through the scales.