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In vivo antibody discovery and hybridoma technology

Published on May 30, 2022   24 min

Other Talks in the Series: The Immune System - Key Concepts and Questions

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0:00
Hi. I'm Katja Hanack, Professor of Immunotechnology at the University of Potsdam in Germany. Today, I'm going to talk about In Vivo Antibody Discovery, especially about Hybridoma technology to generate monoclonal antibodies in vitro.
0:17
To give a short summary about the content. First of all, I will talk about the in vivo process, how antibodies are induced in a mammalian system and in the main part of the talk, I will focus on the technology aspects to induce specific immune responses and how to select feasible binders for several applications.
0:41
On this slide, you can see an overview of how antibody generation is induced in mammalians. In vivo, antibody generation process starts with the entry of a pathogen or a foreign substance, and this entry is recognised by specialised immune cells, such as macrophages or dendritic cells. You see an antigen-presenting cell. These cells are the sentinels of the immune system. They are also called dendritic cells because of their morphology. They have long dendrite by which they can screen their environment for antigens. The cell surface is covered by specialised receptors, so-called pattern recognition receptors, and a famous subgroup is the Toll-like receptors. With those receptors, they are recognising pathogenic structures, such as lipopolysaccharides on bacterial surfaces. Once they have recognised such a structure, they start to internalise the whole pathogen and and degrade it intracellularly in a so-called phagolysosome. In this compartment, we have two different situations. The first one is a very low pH of two, which is degrading pathogenic proteins The second is the appearance of specific enzymes like cathepsin, which help to cleave the protein fragments into smaller peptides (8 to 12 amino acids). These small fragments are loaded on MHC class II complexes and transported to the cell surface. With this process, the dendritic cell is able to present antigens to T cells, and this greenish cell is a T cell, and this T cell has a counterpart receptor, the T cell receptor, by which they can recognise the presented peptide fragment on the MHC complex. This is a speciality of the T cell receptor. The receptor has a dual recognition, it recognises the body's own MHC complex, together with the foreign peptide fragment which is presented by the dendritic cell. This recognition is necessary to activate the T cell and together with other co-stimulatory signals from a co-receptor and different cytokines, the T cell differentiates into an active state. When we have a look at the induction of antibody responses, we need a T helper cell type. This T helper cell type helps the B cell in terms of activation and induction of specific antibody responses.

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In vivo antibody discovery and hybridoma technology

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