Poster

Single-Step Purification Of A 21-mer Oligonucleotide Using Capto™ Q ImpRes Resin

Source: Cytiva
GettyImages-1355122387 RNA

The growing oligonucleotide therapeutics market boasts over 2000 treatments in development and 20 approved drugs, signaling the vast potential of these highly specific sequences. As the pipeline expands, efficient and scalable manufacturing becomes crucial for delivering siRNA and antisense oligo therapies worldwide. Addressing this need, a robust and scalable purification protocol using Capto™ Q ImpRes resin has been developed.

The DNA oligonucleotide synthesis, performed on the ÄKTA oligosynt™ system, achieved a 72% yield with high purity, verified through LC-MS analysis. Impurities, primarily nucleotide losses at the 3’ or 5’ ends, were minimal. Subsequent purification using anion exchange resins demonstrated superior performance with Capto Q ImpRes resin, which offered high dynamic binding capacity (DBC) and purity.

For scale-up, a single-step anion exchange purification on the ÄKTA pilot™ 600 system processed 27 g of crude oligo. The process involved loading a diluted crude oligo solution onto a ReadyToProcess™ column, followed by elution using a gradient of NaOH and NaCl. Fraction pooling based on green recovery strategy achieved >95% purity.

The final product underwent ultrafiltration/diafiltration using a Centramate™ Omega TFF cassette, reaching a concentration of 6.6 mg/mL with 90% recovery and no detectable losses in the permeate. The advancements in synthesis and purification protocols provide a scalable and efficient pathway to meet the growing demand for high-purity oligonucleotide therapeutics.

access the Poster!

Get unlimited access to:

Trend and Thought Leadership Articles
Case Studies & White Papers
Extensive Product Database
Members-Only Premium Content
Welcome Back! Please Log In to Continue. X

Enter your credentials below to log in. Not yet a member of Biosimilar Development? Subscribe today.

Subscribe to Biosimilar Development X

Please enter your email address and create a password to access the full content, Or log in to your account to continue.

or

Subscribe to Biosimilar Development