Question

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

b. What technique should Mr. Ali use 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

The CD4 T-cells are T lymphocytes subtypes that express on the cell surface glycoprotein CD4. For the isolation of CD+T-cells , magnetic based positive or negative selection methods are using. The magcellect human CD4+ T cells isolation kit is designed to human CD4+ T cells from a preperation of peripheral blood mono nuclear cells ( PBMC). Using 2 step procedure that combines both positive and negative selection techniques. The resulting cell preperation is highly enrich for CD4+ T cells with a purity of recovered cells ranging between 85to 95%.

In this technique CD4+ T cells are stained using the conjugated antibodies provided in the kit.