790398 Antibody engineering (in Eng.)

Art
Vorlesung
Semesterstunden
3
Vortragende/r (Mitwirkende/r)
Wozniak-Knopp, Gordana , Traxlmayr, Michael , Wick, Cecilia
Organisation
Biotechnologie und Lebensmittelwissenschaften
Angeboten im Semester
Wintersemester 2024/25
Unterrichts-/ Lehrsprachen
Englisch
Lehrinhalt

The following topics will be covered:

Unit 1

Antibodies, Structures, Domains
Immunoglobulin fold
Mechanism of action, binding Inactivation, effector functions
Diagnostics
Examples of successful antibodies

Unit 2

How to identify new antibodies
Antibody structures,classes, variable and constant domains, hypervariable regions
Hybridomas, mouse antibodies
Humanisation of antibodies
Humanized mouse models

Unit 3
Antibody fragments: Fab, scFv
Designing and making libraries
Randomized oligonucleotides
Parsimonious mutagenesis
Error prone PCR
Library size
Phage Display

Unit 4
Chimeric antigen receptor T cells (CAR T cells)
Architecture of CAR molecules
Biological activities of CAR T cells
Current limitations in the CAR T cell field
Next generation CAR T cells

Unit 5
Yeast Display
Mammalian cell display
Ribosome display
Antigen/antibody interactions
Engineering of stability and activity of antibodies
Mode of action of IgA and IgE antibodies

Unit 6
How to improve existing antibodies and antibody fragments:
Affinity maturations, characterisation of binders, affinity, kinetics
Stabilisation of antibodies
Engineering of in vivo half-life and effector functions

How to characterize antibodies
In vitro assays, receptor binding, cell binding, biological tests with cells (ADCC, CDC, influence on expression of affected genes, ß-Complementation)
In vivo assays, tumor models, rheumatoid arthritis, Immunooncology

Unit 7
Antibody glycosylation and function
Engineering of antibody glycosylation for improved activity

Unit 8
Case report: Engineering the hyperstable protein Sso7d as an alternative binding scaffold
Advantages of small, hyperstable binder scaffolds with Sso7d as an example
Optimization of the Sso7d scaffold for antigen recognition
Engineering Sso7d for specific antigen binding
Exemplary applications of Sso7d-based binders

Unit 9
Side effects of monoclonal antibodies
Developability of antibodies

Unit 10
Alternative antibody formats and bispecific antibodies
scFv fusion proteins
Alternative scaffolds
ADCs

Unit 11
Engineered Abs for treatment of autoimmunity
I-shaped antibodies

Unit 12
Artificial intelligence (AI)-based methods for antibody development
Inhaltliche Voraussetzungen (erwartete Kenntnisse)

The following lectures are highly recommended:

941105 Introduction to Molecular Biology (in Eng.)
772304 Protein chemistry and protein engineering (in Eng.)
791397 Protein engineering (in Eng.)
941319Cell and molecular biology I (in Eng.)
791368 Biological Therapeutics (in Eng.)
Lehrziel

This course gives an overview on current knowledge and methods in antibody engineering, and enables students to choose molecule formats and methods for their discovery and optimization in a problem-oriented manner.

Students who have successfuly completed this course will be able to:
- describe the principles of antibody structure and functions and relate those to engineering
- differentiate between the preferred antibody formats and delineate the optimal solutions for particular applications
- identify the features of these molecules that can be engineered to meet specific requirements, and apply the relevant methods to optimize these properties (e.g. antigen binding or half life in vivo)
- implement the basics of antibody library design and surface display methods, in particular with phage display and yeast display
- differentiate and be able to choose between the various antibody expression platforms, taking into account their advantages and limitations
- apply the acquired knowledge on the methods for biochemical, biophysical and immunological characterization of antibodies and their engineered versions in close-to-reality situations
- elaborate on in vitro and in vivo tools for the characterization of the biological properties of antibodies
- comprehensively describe features of specific groups of antibody-like molecules: alternative binding scaffolds, bispecific antibodies and antibody-drug conjugates
Noch mehr Informationen zur Lehrveranstaltung, wie Termine oder Informationen zu Prüfungen, usw. finden Sie auf der Lehrveranstaltungsseite in BOKUonline.