AIMS AND CONTENT

LEARNING OUTCOMES

"The course of Protein engineering is composed of two parts. The first part is designed to provide students with a basic understanding of the methods used to build expression constructs for the production of modified proteins. The second part describes methods to engineer antibodies used for therapeutic purpose."

AIMS AND LEARNING OUTCOMES

Aim of the course is to provide the student with the basic knowledge on techniques and procedures that constitute the core of protein and antibody engineering. At first, the student will be provided with the basic concepts on recombinant protein production. Afterward, the student will be provided with the different approaches to obtain modified and/or optimized proteins for different applications, with the main techniques used for the aim and with the screening methods.

With respect to antibody engineering the basic concepts on tridimensional immunoglobulin structure and immunoglobulin genetics will be provided. Then, the different techniques available for antibody humanization and the different kind of recombinant antibodies will be discussed.

SYLLABUS/CONTENT

Bacterial espression vectors and bacterial expression systems. Ekaryotic expression vectors and expression systems. Cell-free expression systems. How to choose an eukaryotic expression system. Bicistronic vectors. Vectors for fusion proteins. Vectors for tagging proteins. Inducible expression systems. Alternative cloning techniques. How to build custom vectors. Mammalian expression systems: yeast, insect and mammalian cells.

Immunogloblin structure, immunoglobulin domain, CDR and FR regions, antigen binding site, immunoglobulin constatnt region, basic facts on immunoglobulin genetics. Antiboies used in therapy: from monoclonal antibodies to human antibodies. Monoclonal antibodies, chimeric antibodies, antibody humanization techniques: CDR grafting, resurfacing, superhumanization. Human antibody production using transgenic mice. Immunogenicity of therapeutical antibodies. Alternative recombinant antibodies: scFV, scFV-Fc and minibodies, antibody display platforms, recombinant antibody from camelids (nanobodies).

RECOMMENDED READING/BIBLIOGRAPHY

Suitable books for students are currently not available, slides and references to scientific papers will be provided

TEACHERS AND EXAM BOARD