Description

The growing demand for biotherapeutics and industrial enzymes urge us to progress in the glycosylation of recombinant proteins. Glycosylation is a naturally occurring on the majority of proteins where it contributes to their solubility, stability and biological activity. While the production of recombinant proteins is nowadays performed industrially, their glycosylation remains poorly controlled. The different protein expression systems present specific glycosylation profiles that may differ from those naturally present on the protein. They also generate a large saccharide heterogeneity. A process allowing site-specific glycosylation of any recombinant protein with a precise glycan structure is thus desirable.
The aim of this PhD project is to develop a versatile chemo-enzymatic approach to protein glycosylation by combining synthetic biology and click chemistry. The methodology will be applied to the glycosylation of a therapeutic enzyme used in the treatment of acute lymphoblastic leukemia. This protein will first be produced by bacterial expression system and enzymatically modified in vivo in order to express one or several clickable chemical groups on its surface. Glycosylation will then be performed in vitro with oligosaccharides carrying a clickable chemical group complementary to the one present on the protein. The influence of glycosylation (nature of the oligosaccharide, number and location of the glycosylation sites) on the activity and stability of the protein will be evaluated. The project will be developed within the CBO team (supervision S. Fort) with the support of the microbiology platform supervised by E. Richard.

Skills required

We are looking for a candidate with a strong interest in interdisciplinary projects at the interface between chemistry and biology. He (She) must have advanced knowledge in molecular biology, protein expression, enzymes. Knowledge in organic chemistry or glycochemistry and bioconjugation methods chemistry would be an asset.

Bibliography

Synthetic Glycobiology: Parts, Systems, and Applications, Weston Kightlinger, Katherine F. Warfel, Matthew P. DeLisa, and Michael C. Jewett, ACS Synthetic Biology 2020 9 (7), 1534-1562, DOI: 10.1021/acssynbio.0c00210
Therapeutic enzymes: Discoveries, production and applications, Siddhi Tandon, Anjali Sharma, Shikha Singh, Sumit Sharma, Saurabh Jyoti Sarma, Journal of Drug Delivery Science and Technology 63 (2021) 102455
Bioorthogonal Chemistry and Its Applications, Robert E. Bird, Steven A. Lemmel, Xiang Yu, and Qiongqiong Angela Zhou, Bioconjugate Chemistry 2021 32 (12), 2457-2479, DOI: 10.1021/acs.bioconjchem.1c00461

Keywords

Synthetic Biology, Glycoprotein, Oligosaccharides, Enzyme