Proteins (Major componet in serum. Carriers for other molecules)
Hormones (eg. Insulin and Hydrocortisone- cell attachment and proliferation)
Lipids (eg. Oleic acid) BOUND TO PROTEINS
Minerals (eg. Iron, copper, zinc) BOUND TO PROTEINS
What process is used for making media? What is the material used?
Filtration using a 0.22um filter.
What is the process of producing antiserum?
1) Inject animal with substance of interest (usually mixed with adjuvant)
2) After 1 month, a booster is given
3) Two weeks later, blood is drawn and serum is harvested
What is the difference between the first antigenic challenge and the second antigenic challange?
The second antigenic challenge is:
More immediate in onset
Of a higher titer
Higly specific for that antigen
What is polyclonal antiserum?
Polyclonal antiserum contains antibodies that bind to an antigen, but at different binding sites or affinities. This is because they have differenct hypervariable regions (where they bind to the antigen).
Where do antibodies come from?
B cells that are primed to proliferate turn to plasma cells which make antibodies. Cytokines drive differentiation and proliferation of B cells to become plasma cells.
What are monoclonal antibodies?
Monoclonal antibodies come from the same B cell. They bind to the same antigen at the same site and at the same antigen binding site. (They are all the same!)
What are hybridomas?
A hybridoma is a cell line that produces monoclonal antibodies and is immortal.
(Basically Spleen cells+Myeloma Cells)
What does PEG and HAT stand for and what do they do?
PEG - Polyethylene glycol
fuses myeloma and spleen cells
HAT - Hypoxanthine aminopterin thymidine
Blocks a pathway in myeloma cells so they die. Myeloma cells don't have the enzyme HGPRT while spleen cell does, but spleen cell dies within 2 days.
What are 2 hosts that you can use?
Balb/c mice and rat.
How to choose myeloma cell line?
The cells must not synthesise immunoglobulin, must die in selective HAT media, and must be immortal.