Cation exchange as a single polishing step for conjugated peptides. (4th February 2022)

• Record Type: Journal Article
• Authors: Rockwell, Lauren; Bao, Haiying; Insaidoo, Francis; Ikechukwu, Ijeoma; Tugcu, Nihal; Kandula, Sunitha
• Is Part Of: Biotechnology progress. Volume 38:Number 3(2022)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2022-02-04
• Subjects: conjugated peptide -- ion exchange -- purification -- reverse phase; Biotechnology -- Periodicals; Food industry and trade -- Periodicals; Bioengineering -- Periodicals; 660.6
• DOI: 10.1002/btpr.3238
• Languages: English
• ISSNs: 8756-7938

Abstract

Abstract: Purification of peptides typically includes expensive reverse phase (RP) processes, which utilize high pressure and large volumes of solvent. For two conjugated peptides, chromatography process development targeted a low‐pressure aqueous process that could achieve target product purities of ≥95%, comparable to purities seen with traditional RP. A high throughput screening approach of different modalities was used to identify binding and elution conditions on a cation exchange resin and small‐scale columns were used in order to assess impurity removal and process yield. The parameters for load and gradient elution were optimized to increase product purity and process productivity with a wide operating window identified where high purity and productivity are achieved. Computational modeling was then used to validate experimental chromatography results and to gain insight on the effect of the chemical modifications on the surface properties of the two peptides. Both modeling and experimental data showed that with optimization, cation exchange could be utilized as a single polishing step for conjugated peptides. Similar purities were achieved as those seen with RP with up to double the productivity.