Biotechnology Journal
Biotechnology Journal
5 years ago

Reaction-mediated desorption of macromolecules: novel phenomenon enabling simultaneous reaction and separation

Noriko Yoshimoto, Yuhi Kishi, Aleš Podgornik, Yu Isakari, Shuichi Yamamoto
Combining chemical reaction with separation offers several advantages. In this work we investigated possibility to induce spontaneous desorption of adsorbed macromolecules, once being PEGylated, through adjustment of the reagent composition. Bovine serum albumin (BSA) and activated oligonucleotide, 9T, were used as the test molecules and 20kDa linear activated PEG was used for their PEGylation. BSA solid-phase PEGylation was performed on Sepharose FF Q. Distribution coefficient of BSA and PEG-BSA as a function of NaCl was 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 was adjusted by using NaCl. Spontaneous desorption of PEG - BSA was 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 was applied for activated 9T oligonucleotide, this time using monolithic CIM Q column for adsorption. Selectivity of over 2000 was obtained by proper adjustment of PEG reagent composition. High selectivity enabled spontaneous desorption of PEG-9T without any desorption of activated 9T. Both experiments demonstrated that reaction-mediated desorption of macromolecules is possible when the reaction conditions are properly tuned.

Publisher URL: http://onlinelibrary.wiley.com/resolve/doi

DOI: 10.1002/biot.201700738

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