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(A*STAR) Next generation sequencing methods for glycoconjugates


Project Description

This project builds on a highly successful existing collaboration between the Manchester (existing collaboration between Flitsch/Barran and Field) and A*STAR laboratories (Nguyen-Khuong), including a current A-star studentship (2017-2020 to Ed Pallister). The student will use a combination of synthetic, analytical and computational techniques to develop a next generation sequencing platform for carbohydrates and glycoconjugates. The chemical synthesis of carbohydrates is challenging, but both the Flitsch and Field laboratories have developed enzymatic methodologies, which are easy to perform. The relevant enzymes will be provided from on-going projects. The synthetic analytes will be complemented throughout the project by commercial standards (provided by A*STAR) and at the end by ‘real’ biological samples (e.g. mixtures generated by glycoenzymes, biopharmaceuticals, glycolipids) to test performance.

The second aspect of the project will focus on analytical techniques (existing collaboration between Flitsch and Barran), including chromatography, mass spectrometry and ion mobility spectrometry. Relevant instrumentation is available both in Manchester (where the student will be trained in year one) and at A*STAR, allowing for the seamless transition of know-how and expertise and a cohesive research project. The third aspect of the project will involve computational methods, including applications of databases (some of which exist already) and computational tools to deal with structural identification through complex but unique data fingerprints that will be generated using the methods above. The project will be embedded in current activities both at Manchester and A*STAR .

Entry Requirements:
Applications should be submitted online and candidates should make direct contact with the Manchester supervisor to discuss their application directly. Applicants must have obtained, or be about to obtain, at least an upper second class honours degree (or equivalent) in a relevant subject.

Funding Notes

This project is available to UK/EU candidates. Funding covers fees (UK/EU rate) and stipend for four years. Overseas candidates can apply providing they can pay the difference in fees and are from an eligible country. Candidates will be required to split their time between Manchester and Singapore, as outlined on View Website.

As an equal opportunities institution we welcome applicants from all sections of the community regardless of gender, ethnicity, disability, sexual orientation and transgender status. All appointments are made on merit.

References

[1] Gray CJ, Migas LG, Barran PE, Pagel K, Seeberger PH, Eyers CE, Boons G-J, Pohl NLB, Compagnon I, Widmalm G , and Flitsch SL - Advancing Solutions to the Carbohydrate Sequencing Challenge. Journal of the American Chemical Society, 2019, 141, 37, 14463-14479.
[2] Wongtrakul-Kish K, Walsh I, Sim LC, Mak A, Liau B, Ding V, Hayati N, Wang H, Choo A, Rudd PM, and Nguyen- Khuong T, Combining Glucose Units, m/z, and Collision Cross Section Values: Multiattribute Data for Increased Accuracy in Automated Glycosphingolipid Glycan Identifications and Its Application in Triple Negative Breast Cancer. Anal. Chem. 2019, 91, 9078−9085.
[3] Pallister EG, Choo MSF, Tai J-N, Leong DSZ, Tang W-Q, Ng SK, Huang K, Marchesi A, Both P, Gray C, Rudd PM, Flitsch SL, Nguyen-Khuong, T - Exploiting the Di-Sialyl Galactose Activity of α2,6 Sialyltransferase from Photobacterium Damselae to Generate a Highly Sialylated Recombinant Alpha-1-Antitrypsin. Biochemistry, 2019 in press.
[4] Gray C, Schindler B, Migas LM, Picmanova M, Allouche A, Green A, Mandal S, Motawia M, Sánchez-Pérez R, Bjarnholt N, MøllerB, Rijs A, Barran P, Compagnon I, Eyers C, Flitsch SL, Bottom-up elucidation of glycosidic bond stereochemistry. Anal. Chem. 2017, 89 (8), 4540-4549.
[5] Both P, Green AP, Gray CJ, Sardzik R, Voglmeir J, Fontana C, Austeri M, Rejzek M, Richardson D, Field RA, Widmalm G, Flitsch SL, Eyers CE, Discrimination of epimeric glycans and glycopeptides using IM-MS and its potential for carbohydrate sequencing NATURE CHEMISTRY. 6(1); 65 – 74.

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