Thesis work: Bioorthogonal Labeling of Proteins

Job description
Thesis Project: Bioorthogonal Labeling of Proteins for Biochemical Assays
If you’re inspired by the possibilities of science to make a difference and ready to discover what you can do – join us!

Thesis type: 60 credit
Project start: January 2023
Student location: On Site, Gothenburg Sweden

Background: Biochemical assays are critical in several stages of drug discovery, including high-throughput screening (HTS), lead optimization, and studies of structure-activity-relationship (SAR) and mode-of-action (MoA). A large proportion of drug targets are proteins for which the only known biological function is ligand binding. Such targets require biochemical assays that can measure the interaction between the protein target and a ligand. Often the protein target is expressed as a fusion protein with an affinity tag that can be indirectly “labelled” by a fluorophore-conjugated antibody or other labelled affinity reagent. While this strategy is often successful, it is resource intensive to produce the many protein variants that may need to be tested and there is a significant cost for the labelled affinity reagents. An alternative approach is to directly conjugate a fluorophore to the protein target of interest using a bioorthogonal chemical reaction resulting in a protein target covalently bound to a fluorophore while maintaining its biological activity. Examples of such reactions are amine- or thiol-coupling using N-hydroxysuccinimide- or maleimide-activated fluorophores, respectively. This research is aimed at exploring bioorthogonal protein labelling for biochemical assays and establish a generic labelling protocol for biochemical time-resolved FRET assays.
Hypothesis: Covalently attaching fluorophores to protein targets will result in robust biochemical FRET assays and reduce resource requirements for assay development.
Purpose: Generate fluorophore-conjugated proteins for use in fluorescence applications such as biochemical binding assays to simplify and streamline assay development.

At AstraZeneca, all of our employees make a difference to patients’ lives every day. We operate in more than 100 countries around the world and are one of Sweden’s most important export companies.
 
With more than 2,400 employees from 50 countries, the vibrant Gothenburg site helps to support the entire life-cycle of AstraZeneca medicines, from drug discovery and clinical trials, to global commercialisation and product maintenance. Gothenburg is one of AstraZeneca’s three strategic, global R&D centres, alongside Cambridge and Gaithersburg, and plays a central role in our mission to deliver life-changing medicines to patients.



Responsibilities


Investigate bioorthogonal labelling for a number of protein targets
Optimize labelling reaction conditions and sample purification procedures
Determine the site of labeling by mass spectrometry
Optimize the methodology to achieve site-specific 1:1 labelling
Establish a generic protocol to reproducibly generate fluorophore-conjugated proteins using bioorhtogonal chemical reactions
Develop FRET assays using fluorophore-conjugated proteins and compare to FRET assays that use other affinity reagents such as antibodies






Qualifications
Essential to role:
Strong background in biochemistry
Practical biochemical laboratory experience, including pipetting, handling of solid and liquid chemicals, handling of biomolecules
Good understanding of protein chemistry, including protein purification
Good understanding of biochemical assays and ligand binding kinetics
Good understanding of fluorescence and FRET


Additional desired skills:
Practical experience in biochemical assays and/or protein purification
Practical experience of pipetting small volumes of liquid (<10 µl)
Good understanding of organic chemistry
Good understanding of statistics




About the company
If you’re inspired by the possibilities of science to make a difference and ready to discover what you can do – join us! 

At AstraZeneca we are guided in our work by a strong set of values, and we’re resetting expectations of what a bio-pharmaceutical company can be. By truly following the science, we pioneer new methods, new thinking and bring unexpected teams together. From scientists to sales, lab techs to legal, we’re on a mission to turn ideas into life-changing medicines that transform lives. We need great people who share our passion for science and have the drive and determination to meet the unmet needs of patients around the world. If you’re swift to action, confident to lead, willing to collaborate, and curious about what science can do, then you’re our kind of person.

AstraZeneca is an equal opportunity employer. AstraZeneca will consider all qualified applicants for employment without discrimination on grounds of disability, sex or sexual orientation, pregnancy or maternity leave status, race or national or ethnic origin, age, religion or belief, gender identity or re-assignment, marital or civil partnership status, protected veteran status (if applicable) or any other characteristic protected by law. AstraZeneca only employs individuals with the right to work in the country/ies where the role is advertised.