Cation exchange as a single polishing step for conjugated peptides

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%, co...

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Bibliographic Details
Published inBiotechnology progress Vol. 38; no. 3; pp. e3238 - n/a
Main Authors Rockwell, Lauren, Bao, Haiying, Insaidoo, Francis, Ikechukwu, Ijeoma, Tugcu, Nihal, Kandula, Sunitha
Format Journal Article
LanguageEnglish
Published Hoboken, USA John Wiley & Sons, Inc 01.05.2022
Wiley Subscription Services, Inc
Subjects
Cation exchange
Cation exchanging
Cationic polymerization
Chromatography
Computer applications
conjugated peptide
Elution
High pressure
High-throughput screening
ion exchange
Ions
Optimization
Peptides
Polishing
Process parameters
Productivity
Protein purification
purification
Purity
reverse phase
Surface properties
conjugated peptide
reverse phase
purification
ion exchange
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Summary: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.
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ISSN:8756-7938
1520-6033
DOI:10.1002/btpr.3238