BRAIN-STORMING ASSIGNMENT –

The gonadotrophic hormones luteinizing hormone (LH), follicle stimulating hormone (FSH), and human chorionic gonadotropic hormone (HCG, produced during pregnancy) and thyroid stimulating hormone (TSH) share general structural characteristics. All are glycoproteins composed of two peptide subunits, an alpha and a beta protein chain. The alpha subunit is identical in all four hormones. The beta subunit is specific for each hormone and is responsible for their unique biological activities. However, the difference between LH and HCG are small regarding functional abilities and amino acid sequence of the beta subunit; only the 36 aa carboxyterminal portion (aa 109-145) of the HCG beta chain differs from the LH beta chain.

HCG is produced by trophoblast cells (those cells of the embryo which will form the placenta) already about six days after fertilization. The presence of HCG in the maternal urine and plasma is the first objective sign of an established pregnancy.

A synthetic carboxyterminal peptide of HCG (aa 109-145) is commercially available but is by itself not very immunogenic and needs to be conjugated to a protein carrier (e.g., diptheria toxoid) prior to immunization.

Design a "use at home" pregnancy kit

Hints: antibody has to be specific for HCG and may not cross react with LH.

A women will not have a fluorescent microscope in her home. Since the product is sold in the drugstore it should not require refrigeration and should not contain labile or dangerous reagents.

Not all antibodies in the system have to be prepared by you nor do they have to be monoclonal – you can buy them

Address the following:

To get HCG specific monoclonal antibody

- immunization protocol

- fusion

- screening assay

• final test kit with necessary controls – complete with materials and procedure

To put together the pregnancy testing kit

• write out and draw steps how you use the necessary antibodies and how reagents interact with each other, how do you get a ‘yes or no’ answer

• write out steps on what the woman has to do, think through the complete process

You need to pass in a copy of your notes after the brain storming session and then we go through “a design effort” together.

Group 2

Design a vaccine for the virus Porcine mortales (a virus of pigs) based on monoclonal antibodies. The virus is lethal to pigs if injected alive. Killed virus or any fraction thereof is not immunogenic in pigs. The live virus is however not lethal in mice and is immunogenic.

Discuss: immunization protocol

fusion

screening assay

vaccine testing

Ref.: Dr. Gisela Erf. University of Arkansas