• Created by Unknown User (lylaatta@mit.edu), last modified by Unknown User (clrichar@mit.edu) on Jul 13, 2015 15:23

Progress

CloningTransfectionInductionCytometryData Analysis
     

 

Procedure

Notes: While we can determine how level of induction will affect receptor signal to noise ratio, we will not be able to determine the concentration of the receptor in the cell without pairing it to a fluorescent reporter which would take too much time and money to construct.

Optimal receptor vs Syk-TEV concentration is likely to vary for each antigen detected by a different antibody as the different antigens have different Kd values.

      
      
      
      
      
      
      
      

Background

Once we have optimized the CD79A and CD79B linkers in our receptor, the expression levels of both the receptor and Syk-TEV need to be optimized. Since there is a relatively high level of background cleavage of the TCS by ambient Syk-TEV, it is likely that extremely low expression of Syk-TEV will give us the best signal to noise ratio. 

Approach

Because both receptor concentration and syk-TEV concentration will affect eachother we must treat this as a 2D optimization problem. We will form a 6x6 grid with varying expression of the receptor across one axis and varying expression of Syk-TEV across the other. We will then pick the point with the best signal to noise ratio to continue experimentation.

Parts

Results

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