Academy Project: Investigating feasibility of scaling up protein purification of a membrane protein

Aquaporin A/S is a pioneering biotechnology company at the forefront of water purification and filtration. By harnessing the natural water channels present in living organisms, Aquaporin specializes in the development of cutting-edge membrane technologies that facilitate highly efficient water treatment processes. At the heart of its innovation lies the Deep Tech Center, where research is dedicated to the advancement of the next generation of biomimetic membranes. This next phase focuses on every aspect of biomimetic membrane development, from protein production to protein stabilization and integration into the filtration membrane. Aquaporin's commitment to pushing the boundaries of biomimicry promises to revolutionize water purification, offering sustainable solutions to address pressing global water challenges.

Objective
The primary objective of this project is to assess the viability of scaling up the purification process for one to six variants of a membrane protein. Beginning with an established academic protocol, the project will transition into a production protocol necessitating adjustments in volume, chemicals, equipment, and other factors. Post-purification, the focus will shift towards investigating the parameters required to stabilize the various proteins into a crystalline 2D sheet. This endeavor aims to bridge the gap between academic research and industrial application, paving the way for potential advancements in protein purification and stabilization techniques.

Tasks
The project encompasses a comprehensive set of tasks, blending literature study with experimental procedures to explore membrane proteins, membrane protein purification, and 2D sheet formation. The experimental tasks include:

• Affinity Chromatography: Employing this technique to isolate specific membrane proteins from complex mixtures, leveraging their unique binding properties to affinity ligands.
• SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis): Separating proteins based on molecular weight using an SDS denaturing agent to unfold and standardize protein charges.
• Western Blot: Detecting and characterizing specific proteins within a complex sample using antibodies that bind to target proteins separated by SDS-PAGE.
• Native-PAGE: Separating proteins in their native state without denaturation, allowing for analysis of protein-protein interactions and native protein conformations.
• HPLC-SEC (High-Performance Liquid Chromatography Size-Exclusion Chromatography): Utilizing this technique to separate proteins based on size, shape, and hydrodynamic volume.
• BCA Assay (Bicinchoninic Acid Assay): Quantifying protein concentration in samples through colorimetric detection of copper ions reduced by protein molecules.
• 2D Sheet Formation: Investigating the process of stabilizing purified proteins into crystalline 2D sheets.

Student Opportunities and Profile
The project represents a blend of theoretical exploration through a literature study and practical application via hands-on experimentation, providing the student with a comprehensive learning experience. Beginning with an in-depth literature review, the student will delve into the existing research on membrane proteins, membrane protein purification, and 2D sheet formation, gaining a solid theoretical foundation. Subsequently, the student will undergo thorough training to develop the necessary laboratory skills and techniques. With this preparation, the student will have the opportunity to independently conduct experiments in the laboratory, applying his/her knowledge to tasks such as affinity chromatography, SDS-PAGE, Western blotting, and 2D sheet formation. This hands-on approach fosters critical thinking, problem-solving, and technical proficiency, empowering students to contribute meaningfully to the project while gaining valuable research experience in a real-world laboratory setting. Ideal candidates for this opportunity possess a strong interest in biotechnology or biochemistry, a passion for scientific inquiry, and a willingness to learn and explore new concepts and techniques.

Project Type

• Internship
• Special course
• Bachelo'rs Thesis, or
• Master's Thesis

Project timing
Autumn semester 2024, for a minimum of 3 months preferably 6 months, depending on the type of project.

All work will be conducted in a safe manner according to Danish safety standards.

Application deadline:
15. May 2024