Questions

Question 1

How might you create a stable cell line?

Answer 1

Long-term expression

6-8 months to make

Same complex for getting plasmid in
Lipo- or electro-
Antibiotic selection
Dilution to single cells

ELISA 
MTX amplification
Select for most productive cells 
Binding amount 
Serial dilution across the plate
Drop off in production with dilution
Still high production at low dilution = high producers 

Foreign gene incorporated into mammalian genome
Same complex for getting plasmid in
Lipo- or electro-

Selectable marker
Essential cell component deficient
E.g. Di-hydrofolate reductase deficient
Recombinant plasmid contain gene for di-hydrofolate reductase

Question 2

Describe how you may express fully humanised antibodies

Answer 2

Phage display

Ribosome display

Question 3

Describe how you may express fully humanised antibodies

Answer 3

Phage display

Ribosome display

Question 4

What is the future for antibody therapeutics?

Answer 4

Next Generation antibody therapeutics

Antibody Drug conjugates
Specific monoclonal antibody with highly potent cytotoxic agents attached by biodegradable linker
To deliver cytotoxic agent(s)
Why use an antibody?
Increases specificity of targeting 

Bispecifics
One antibody, two distinct binding arms
Bind to different antigens or epitopes on the same antigen
E.g. retarget T cells to kill tumour cells
1 - binds to cancer cell
2 - binds to recruit cytotoxic T cell

CAR-T therapy PERSONALISED MEDICINE
Chimeric antigen receptor - T (cell) therapy
Type of adoptive cell therapy (ACT)
Chimeric antigen receptor - antibody
Engineer T cells from patients
T cells clear tumour cells/cancer cells
Make antibody to help T cells to find cancer cells
Insert antibody gene into T cell
T cells now express chimeric antibody on the surface - bind to specific targets on tumour cells
Put modified T cells back into patient

Question 5

What is the future for antibody therapeutics?

Answer 5

Next Generation antibody therapeutics

Antibody Drug conjugates
Specific monoclonal antibody with highly potent cytotoxic agents attached by biodegradable linker
To deliver cytotoxic agent(s)
Why use an antibody?
Increases specificity of targeting 

Bispecifics
One antibody, two distinct binding arms
Bind to different antigens or epitopes on the same antigen
E.g. retarget T cells to kill tumour cells
1 - binds to cancer cell
2 - binds to recruit cytotoxic T cell

CAR-T therapy PERSONALISED MEDICINE
Chimeric antigen receptor - T (cell) therapy
Type of adoptive cell therapy (ACT)
Chimeric antigen receptor - antibody
Engineer T cells from patients
T cells clear tumour cells/cancer cells
Make antibody to help T cells to find cancer cells
Insert antibody gene into T cell
T cells now express chimeric antibody on the surface - bind to specific targets on tumour cells
Put modified T cells back into patient

Question 6

What is the future for antibody therapeutics?

Answer 6

Next Generation antibody therapeutics

Antibody Drug conjugates
Specific monoclonal antibody with highly potent cytotoxic agents attached by biodegradable linker
To deliver cytotoxic agent(s)
Why use an antibody?
Increases specificity of targeting 

Bispecifics
One antibody, two distinct binding arms
Bind to different antigens or epitopes on the same antigen
E.g. retarget T cells to kill tumour cells
1 - binds to cancer cell
2 - binds to recruit cytotoxic T cell

CAR-T therapy PERSONALISED MEDICINE
Chimeric antigen receptor - T (cell) therapy
Type of adoptive cell therapy (ACT)
Chimeric antigen receptor - antibody
Engineer T cells from patients
T cells clear tumour cells/cancer cells
Make antibody to help T cells to find cancer cells
Insert antibody gene into T cell
T cells now express chimeric antibody on the surface - bind to specific targets on tumour cells
Put modified T cells back into patient