Question

Mr. X collected blood samples from different subjects and he is planning to isolate CD4+ T-cells only. After the isolation of CD4+ T-cells he wants to extract protein and study the expression of CD4 (which is a glycoprotein) among the subjects. Based on these, answer the following:

a. What technique should Mr. X use to isolate CD4+ T-cells from other blood cells? Give a brief description for this technique

b. What technique should Mr. Xuse to separate the protein he interested in?

c. State the main steps and briefly the science behind each step of the technique that you answered in question b

(150-250 words)

Answer 1

a) Cell seperation technique based on antibody-magnetic beads can be used to isolate CD4+T cells from other blood cells.

In cel suspension , antibody-magnetic beads are added .These antibody magnetic beads are precoated with target specific antibodies here antibodies against CD4+ T cells. The CD4+T cells will get attached to the antibody-magnetic beads and thus a magnet is used to select the target cells (positive selection ) and the other cells won't attached to the magnet as it has no antibody-magnetic beads  ( negative selection). Then these magnetic beads are deattached by a polyclonal antibody that binds to the Fab region of the specific monoclonal antibody ,thereby alternating its affinty fro the antigen, The isolated cells are left without antibody in their surface.

b) Affinity chromatography can be used to seperate the protein of interest.

c)