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| Author(s): |
Caio Cesar Nogueira Cambui
Total Authors: 1
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| Document type: | Doctoral Thesis |
| Press: | São Paulo. |
| Institution: | Universidade de São Paulo (USP). Conjunto das Químicas (IQ e FCF) (CQ/DBDCQ) |
| Defense date: | 2023-08-21 |
| Examining board members: |
Ricardo José Giordano;
Daniela Sanchez Bassères;
Nathalie Cella
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| Advisor: | Ricardo José Giordano |
| Abstract | |
Antibodies are biomolecules used by the immune system to identify and neutralize foreign bodies such as bacteria, viruses and/or tumor cells. Monoclonal antibodies have a broad range of applications, being used as scientific tools and even as biodrugs to tread a variety of diseases. However, the production of antibodies is challenging. The main techniques used in this process are the hybridoma technology and antibody phage display. These techniques, however, rely on animal immunization, which is a limiting factor, especially when the antigen used is toxic, non-immunogenic or a self-antigen. In this study, we combined peptide and antibody phage display to develop an alternative method to generate antibodies. Peptide phage display allows the isolation of bioactive peptides. These peptides can be engineered in an antibody scaffold, allowing the construction of libraries directed against any target. Therefore, these libraries allow the isolation of immunoglobulins that bind biological active sites located on the antigen. We demonstrated the viability of this process by isolating antibodies against Tie1. Tie1 is a tyrosine receptor kinase (RTK) expressed by endothelial cells. This receptor plays a key role in angiogenesis, the formation of new blood vessels from pre-existing ones. Tie1 is a member of a family of RTKs composed by only two members: Tie1 and Tie2. Tie2 is an extensively studied receptor. Its ligands and pathways are well known. However, little is known about Tie1, which stills an orphan receptor. We isolated a peptide directed against the extracellular region of Tie1 using peptide phage display. This peptide displayed an antiangiogenic activity on a murine model of oxygen-induced retinopathy and was used to construct an antibody library directed against Tie1. The isolation of specific single-chain antibodies against Tie1 demonstrates the efficiency of our method. As a second part of this work, we isolated a peptide directed against the extracellular portion of gp130, a pleiotropic glycoprotein receptor that plays important roles in several biological processes, such as inflammation andangiogenesis. We hope that this peptide may be used to investigate pathways related to the immune response, inflammation and angiogenesis. (AU) | |
| FAPESP's process: | 19/19620-5 - Peptide and antibody phage display: a combination for Engineering antibodies against angiogenesis dependent diseases |
| Grantee: | Caio Cesar Nogueira Cambui |
| Support Opportunities: | Scholarships in Brazil - Doctorate (Direct) |