Pfizer, Inc., 700 Chesterfield Village Parkway, Chesterfield, MO 63017, USA.
J Chromatogr A. 2010 Jun 11;1217(24):3783-93. doi: 10.1016/j.chroma.2010.04.019. Epub 2010 Apr 24.
A cation exchange chromatographic purification process step was developed for the purification of human PEGylated PYY 3-36 from the PEGylation reaction mixture. In this publication we describe experiments carried out to evaluate the chromatographic performance of arginine chloride as an effective cation exchange chromatography eluent. Using arginine we obtained improved recovery and resolution during chromatographic purification of a peptide PEGylation reaction mixture. The chromatographic elution performance of arginine was compared to other cationic amino acids and sodium chloride. Arginine provided higher yield and better resolution of product from other process impurities. The process was successfully scaled up to produce clinical supplies. The basis for improvement in process performance with arginine was characterized by examining the effect of buffer and concentration of the PEGylated peptide on hydrodynamic volume of the molecule in solution. These results were used to predict the behavior of the molecule in the chromatography process. The enhanced chromatographic performance could be attributed to changes in molecular size with concentration, higher eluent strength of arginine, and resulting changes in mass transfer resistance.
建立了一种阳离子交换色谱纯化工艺步骤，用于从 PEG 化反应混合物中纯化人 PEG 化 PYY3-36。在本出版物中，我们描述了进行的实验，以评估盐酸精氨酸作为有效阳离子交换色谱洗脱剂的色谱性能。使用精氨酸，我们在色谱纯化肽 PEG 化反应混合物时获得了更高的回收率和分辨率。将精氨酸的色谱洗脱性能与其他阳离子氨基酸和氯化钠进行了比较。与其他工艺杂质相比，精氨酸提供了更高的产物收率和更好的分辨率。该工艺已成功放大生产临床供应品。通过考察缓冲液和 PEG 化肽浓度对溶液中分子流体力学体积的影响，确定了精氨酸改善工艺性能的基础。这些结果用于预测分子在色谱过程中的行为。增强的色谱性能可归因于浓度下分子大小的变化、精氨酸更强的洗脱能力以及传质阻力的变化。
