Research project BL/02/SA14 (Research action BL)
Aim :
This trans-disciplinary network (chemistry & immunology) will engineer an immunosensor platform for real time signalling of biomarkers for priority diseases such as Tuberculosis (TB) (https://bccm.belspo.be/), breast cancer (BC) and Covid-19, by the integration of replaceable target-specific nanobody reactor/chip, electrochemiluminescent (ECL) signal transduction and microfluidic technology. This network will leverage submission of joint projects https://www.ugent.be/nl/onderzoek/financiering/bof/dos/overzicht.htm for students of UWC to apply for a PhD fellowship at Ghent university.
Objectives
(i) Preparation/generation of target-specific nanobodies of biomarker (antigens): interferon gamma (IFN), human epidermal growth factor receptor 2 (HER2) and SARS-CoV-2 proteins spike (S), nucleocapsid (N) and envelop (E), for TB, BC and Covid-19, respectively.
(ii) Preparation of nanobody-dye or nanobody-quantum dot (QD) conjugates. (dye = tris(2,2’- bipyridyl)ruthenium(II) [Ru(bpy)3 2+] luminophore).
(iii) Preparation of sandwiched ECL immunosensing reactor with capture (primary nanobody) and signalling antibody (dye-tagged or QD-tagged secondary nanobody) on indium tin oxide (ITO) and Au wafer electrodes or boron-doped diamond microparticles.
(iv) Integration of microfluidics device with the immunosensing ECL reactor to form the sensor platform for biomarkers.
(v) Studying the response dynamics of the fabricated sandwich ECL immunosensor.
Specific R&D cooperation topic(s)
Objectives (i) and (ii) performed at Ghent University (GU) by 2 PhD students, while Objective (iii) – (v) will be done at the University of the Western Cape (UWC). PhD student (Miss Jaymi January) and early career researcher (Dr Keagan Pokpas) would visit GU to work on conjugation of the secondary nanobody. GU early career researcher would participate in Objective (v) through validation studies using ELISA.
Quality, excellence of individual network members
Prof Dr Gettemans (PI at GU) is at the Department of Biomolecular Medicine and he heads the Nanobody Lab. He has expertise on production of antigens, development of target-specific nanobodies and immunochemistry. The Nanobody Lab has the facilities required for the project. Prof Emmanuel Iwuoha is the South African Research Chair for Nanoelectrochemistry and Sensor Technology. He has expertise on various types of new generation electrochemical biosensors for disease biomarkers. The work being proposed will be done at UWC SensorLab which consists of well-equipped laboratories for electrochemistry studies. Dr Keagan Pokpas is an early career lecturer at UWC with experience in electrochemical sensors. He has grants from the South Africa Medical Research Council (SAMRC) and National Research Foundation (NRF) and has publications in the area of electro-bioanalysis. Jaymi January is a female PhD student. She has a master’s level experience in the development of biosensors.
Expected Results
i. New nanobody libraries for interferon gamma (IFN), (HER2) and SARS-CoV-2 proteins.
ii. Protocol will be developed for the conjugation of specific nanobodies with luminophore probes (dyes and QDs).
iii. Novel sensing platform technology that functions by replacement of used reagents within the reactor and with reagents of the nanobody of the target antigen (biomarker) will be developed for.
iv. Training and graduation of 1 MSc and 1 PhD student from UWC and on nanobody immunochemistry and nanobody immunosensor technology. v. Training 1 PhD student from GU (Faith Njeri) on immunosensor technology.
vi. Co-authoring of publications, conference presentation and patents.
