Class: Crash Course Genetech Oral Exam (additional requirement for MSc Students)
Date: WS 2023

Q1: He starts off very general: "You find a gene associated with a protein. What do you do with it to find a treatment?"
A1: He wants you to get to gene sequencing first and then you will eventually talk all the way through DECL and antibody-phage display libraries with the exam. So first, talk about gene sequencing of the diseased gene, comparing it to a healthy gene to look for differences. Maybe also mention transcriptomic methods like q-RT-PCR where you can compare relative protein quantification differences and that that may indicate cancer/diseased state.

Q2: You have the gene and now you want to express the protein. What do you do?
A2: PCR (explain ingredients and steps), explain vector insertion (mention ligation, restriction enzymes, restriction sites on the plasmid and DNA) (he asked a follow up question about types of vectors and their differences then asked me to keep explaining), when vector is constructed you insert into the cell (how? - electroporation), then grow the cell (in what?), select cell (how? explain antibiotic resistant genes in vector, maybe even name some examples), lysing cell and wash to keep just protein

Q3: How do you make monoclonal antibodies?
A3: antigen transfect mouse to get immune response, remove spleen cells (you want B cells), fuse with immortal cancer cells aka. myeloma cells to get hybridomas (immortal), plate and grow, select grown antibodies

Q4: What is another way to make antibodies?
A4: antibody-phage display libraries, then talk about it and explain whole process of how it works (panning 3x, affinity maturation)

Q5: how can you make antibodies more specific?
A5: talk about hypervariable regions (CD3, CD2, CD1), process of introducing genetic variation

Q6: do you know about other types of display libraries?
A6: DECL - explain whole process (know this stuff well because this is his area of research)

Q7: small molecules vs. macromolecules as drugs - advantages and disadvantages?
A7: small molecules have better tumor perfusion than macromolecules but are less specific

He will ask a lot of follow up questions to get you to really walk him through the whole process, with as much detail as you can give him. He cares about vocab a lot and will ask questions beyond what you learned in the course if you show him that you know a topic well (e.g., he asked me what other display libraries I knew after talking about DEL and antibody-phage display - I only mentioned that Yeast-based display libraries exist but did not have to go into detail).

You can definitely steer him a bit and take your time with details on topics you know well.