790398 Antibody engineering (in Eng.)


Art
Vorlesung
Semesterstunden
3
Vortragende/r (Mitwirkende/r)
Steinkellner, Herta , Wozniak-Knopp, Gordana , Traxlmayr, Michael , Rüker, Florian
Organisation
Angeboten im Semester
Wintersemester 2021/22
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

Yeast Display
Ribosome Display

Unit 5

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 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

Unit 7

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 8

Expression systems for antibodies and antibody fragments
Antibody expression systems (advantages and limitations)
E.coli, yeast, insect cells, mammalian cells, plant (cells)

Unit 9
Antibdy glycosylation and function
Antibody engineering of posttranslational modifications
Challenges in the development of SARS-CoV-2 antibodies

Unit 10

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

Unit 11

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 12

Case report: Modular antibodies - Engineering antigen binding sites into the Fc region of IgG

Exam

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, functions, 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.
Noch mehr Informationen zur Lehrveranstaltung, wie Termine oder Informationen zu Prüfungen, usw. finden Sie auf der Lehrveranstaltungsseite in BOKUonline.