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Production and characterization of recombinant antibodies (scFv and bispecific antibodies) against ion channels with potential application for cancer diagnostic and immunotherapy
Universita degli studi di Firenze
Experimental and Clinical Medicine Department (General Pathology Section)
Type of Research Project
- Basic science
What is the background of the project?
The present internship will take place in a cancer biology laboratory, within a multidisciplinary context, dealing with different techniques for the study of cancer behaviour (e.g. molecular biology, tumor cell culture, electrophysiology, in vivo studies, in vivo imaging, antibody production and engineering).In particular the internship will focus on antibody production and engineering against ion channel, with attention to hERG1 ion channel and its related macromolecular complexes. It is known that among hindrances in cancer treatment, the lack of appropriate markers to be exploited for targeted therapy, accompanied by the need of new potential drug represent two of the main challenges. It has been demonstrated that hERG1 potassium channels could represent a novel class of oncological targets, as they are expressed in a variety of tumors of different histogenesis. Moreover, hERG1, inside a hERG1/ẞ1/PI3K complex, sustains angiogenesis and IFMSA International Secretariat, c/o IMCC, Nørre Allé 14, 2200 København N., Denmark progression in colorectal (CRC) and pancreatic cancer (PDAC). In preclinical models, the treatment with classical hERG1-specific blockers overcome chemoresistance in Leukemia and inhibits tumor growth, angiogenesis and metastatic spread in CRC and PDAC. One of the most intriguing aspects of hERG1 channels involvement in tumor establishment and progression is their interaction with adhesion molecules, such as integrins.
What is the aim of the project?
The aim is to characterize and develop antibodies against such complex and other associated channel and transporter (e.g. CAIX, NHE1) which are key regulators in such cross-talk.
What techniques and methods are used?
The above mentioned antibodies will be developed applying molecular biology techniques, such as PCR, affinity chromatography, cloning, bacterial cells manipulation, transfection, tumor cell culture, antibody labelling with fluorophores.
What is the role of the student?
- The student will mainly observe
- The student will observe the practical experiments but will be highly involved in the analysis of the results
- If the project includes “lab work”
- the student will take active part in the practical aspect of the project
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
The student will be actively involved in the development of engineered antibodies, performing cloning and bacterial cell transfection in order to assemble the antibody construct. He/She will be trained in cell culture principal manipulation and also for the use of mammalian and yeast cells for protein expression. He will also be involved in the test of such antibodies on tumor cells of different histogenesis to determine their effects on viability, proliferation, migration. The intern will also acquire more complex techniques, such as IF (immunofluorescence), antibody labelling with fluorophores and cell-ELISA.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
It is required a general knowledge of the basic techniques applied for laboratory research and a broad knowledge of good laboratory practice (GLP). The rest of the teaching will be provided by the tutor during the course of the internship.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- No specific outcome is expected
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Good level of English, to be able to read scientific papers. Laboratory expertise will be acquired on site.
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students
- - Arcangeli A.; Duranti C. Ion channel Targeting with Antibodies and Antibody Fragments for Cancer Diagnosis. Review 2019
- - Duranti C; Carraresi L; Sette A; Stefanini M; Lottini T; Crescioli S; Crociani O; Iamele L; De Jonge H; Gherardi E; Arcangeli A. Generation and Characterization of novel recombinant anti-hERG1 scFv antibodies for cancer molecular imaging. Oncotarget; 2018; vol. 9; (No. 79); pp: 34972-34989
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