Posted: September 5th, 2015

Infection and Immunity

Infection and Immunity – Experimental Design

The maximum length of your answer to this assignment is 5 pages (1.5 spacing – Times New Roman font – 12 point size; excluding bibliography). Any pages additional to this limit will not be marked.

You are a research assistant working in an immunology laboratory. You are provided with (i) 50 mg of a conjugate, comprised of a short 6-mer peptide (6pep) covalently attached to bovine serum albumin (6pep-BSA), (ii) 50 mg of a second conjugate, comprised of 6pep covalently attached to keyhole limpet haemocyanin (6pep-KLH), (iii) 50 mg of BSA, and (iv) 50 mg of KLH. All proteins and conjugates provided to you are highly purified. You are asked to produce several mg of a purified rabbit polyclonal antibody that reacts specifically with 6pep but not with either BSA or KLH. You can access any other items that might be required (e.g. rabbits, secondary antibodies, etc.).

You may or may not need to use all the reagents provided (that is for you to decide).

Like most small molecules, by itself, 6pep is poorly immunogenic and binds poorly to ELISA plates.

Using a flow chart, design an experimental protocol to achieve this. Consider carefully the route and details of immunization, and the number of animals to be used. Comment on the likely specificity of the final product. Relevant information will be provided to you (i) in lectures, (ii) in books held in Closed Reserve in the library, and (iii) by you asking about anything not available via (i) or (ii).

The written assignment requires you to produce an experimental design.

On one page produce a flow chart showing details of the planned experimental strategy. On the remaining 4 pages:

(i) Justify the number of animals used, their post-experimental fate, and the route and details of immunization chosen.
(ii) Briefly describe any special techniques used in the proposed protocol (cite references).

Provide an appropriately formatted bibliography of any references used in completing the assignment (this is NOT included in the 5 page limit; be sure you have cited these references at relevant points in the text of the assignment).

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