PEGylation of therapeutic proteins has been employed to improve biopharmaceuticals clinical performance (considering immunogenicity and plasma half-life). Despite the advantages of PEGylated biopharmaceuticals, the parallel reactions between reactive PEG and the functional amino acids of proteins result in heterogeneity of PEGylated species. This problem associated with the hydrolysis of reactive PEG is limitations of this technique. In addition, the purification of the PEGylated proteins is complex and inefficient, compromising the economical viability of the process. In this project the study of more efficient PEGylation reactions for the model proteins L-asparaginase and catalase using microfluidic technology is proposed. The continuous PEGylation reaction process will be developed in micro-reactors and aqueous biphasic systems (ABS) will be investigated as purification technology, both by microfluidics and by the centrifugal partition chromatography (CPC) platforms. As expected results, it is intended to develop a site-specific PEGylation reaction in micro-reactors for the two therapeutic proteins in a continuous flow reactive media, with higher yield, shorter reaction time and applying less amount of reactive PEG than is currently practiced on a laboratory scale (batch process). Another expected result will be the development of a continuous purification process with ABS associated with the microfluidic reaction stage. Optimization of the continuous purification technique using CPC technology will also be implemented, allowing the efficient separation of PEGylated protein forms with high purity.
News published in Agência FAPESP Newsletter about the scholarship: