M15. 15: Using some commercially available nanoproducts. See: cytodiagnostics.co

Question

M15.

15: Using some commercially available nanoproducts.

See:      cytodiagnostics.com   website,

http://www.lifetechnologies.com/us/en/home/references/molecular-probes-the-handbook/ultrasensitive-detection-technology/qdot-nanocrystal-technology.html

Describe how you would solve the following problem using nanomaterials:

You grow some tumor cells in culture, and you’ve isolated and purified a surface antigen (protein on the cell membrane) that appears to be specific to this type of tumor cell.   You want a fluorescent label to stain these tumor cells in a heterogeneous tissue biopsy. You decide a fluorescent nanocrystal that emitted green light (say 560 nm, excited with a blue laser or LED) would make a good reporter that would selectively bind to the tumor cells in various samples.    This problem involves two stages: 1. Raising specific antibodies against a target protein of interest (use your general biotech knowledge here), and 2. Developing a specific nanoscale reporter for that cell/protein (use your nano knowledge and know-how on these websites).  

Explanation / Answer

Firstly, we need to clone that particular tumor gene in cloning vector and then need to express it in a suitable expression vector followed by transformation into a suitable host and culturing the host cells and purifying that particular protein using affinity chromatography techniques. Now, purified protein is to be coupled with adjuvants for antibody production in an appropriate animal model. Again, need to give booster dose and finally collect the serum for isolation of monoclonal antibody specific to that particular tumor protein.

Now, our interested nanocrystals organic surface layer must be coated with fluorescent dyes (monochromic) coupled specific antibody for that tumor antigen using quantum dot technology. Here, the affinity linker (Ab) coupled to the amphiphilic polymer coating by means of functionalised polyethylene glycol (PEG). It can be useful in live cell imaging and cellular dynamics of tumor by detecting the emitted fluorescence from that antigen-specific tumor cells. It would be necessary to take precautions about size (~20 nm) by altering the core (semiconductor material) composition or density.

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