Reaction‐Mediated Desorption of Macromolecules: Novel Phenomenon Enabling Simultaneous Reaction and Separation. Issue 7 (26th February 2018)

• Record Type: Journal Article
• Authors: Isakari, Yu; Kishi, Yuhi; Yoshimoto, Noriko; Yamamoto, Shuichi; Podgornik, Aleš
• Is Part Of: Biotechnology journal. Volume 13:Issue 7(2018)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2018-02-26
• Subjects: reaction‐mediated desorption -- proteins -- oligonucleotides -- PEGylation -- ion‐exchange chromatography; Biotechnology -- Periodicals; 660.605
• DOI: 10.1002/biot.201700738
• Languages: English
• ISSNs: 1860-6768

Abstract

Abstract : Combining chemical reaction with separation offers several advantages. In this work possibility to induce spontaneous desorption of adsorbed macromolecules, once being PEGylated, through adjustment of the reagent composition is investigated. Bovine serum albumin (BSA) and activated oligonucleotide, 9T, are used as the test molecules and 20 kDa linear activated PEG is used for their PEGylation. BSA solid‐phase PEGylation is performed on Q Sepharose HP. Distribution coefficient of BSA and PEG‐BSA as a function of NaCl is determined using linear gradient elution (LGE) experiments and Yamamoto model. According to the distribution coefficient the selectivity between BSA and PEG − BSA of around 15 is adjusted by using NaCl. Spontaneous desorption of PEG − BSA is detected with no presence of BSA. However, due to a rather low selectivity, also desorption of BSA occurred at high elution volume. A similar procedure is applied for activated 9T oligonucleotide, this time using monolithic CIM QA disk monolithic column for adsorption. Selectivity of over 2000 is obtained by proper adjustment of PEG reagent composition. High selectivity enables spontaneous desorption of PEG‐9T without any desorption of activated 9T. Both experiments demonstrates that reaction‐mediated desorption of macromolecules is possible when the reaction conditions are properly tuned. Abstract : Molecules (green) adsorbed on the surface of the matrix are being solid‐phase PEGylated by activated PEG (red). Due to weakening of interaction (shift of distribution coefficient) after PEGylation, PEGylated molecules are spontaneously desorbed from the surface while original molecules remain adsorbed – phenomenon called reaction‐mediated desorption of macromolecules. Blue arrow indicates molecule that is just PEGylated and it is in the process of desorption.