Biotin/Biotin-HRP Competitive Binding - Report Writing Science Assignment Help

Assignment Task:

Some key points for writing the ELISA report: 

Don’t focus just on the method. You are using the ELISA technique to solve a problem.  Remember the problem comes from an accident at a Biotech company and you need to  determine if biotinylated proteins are present (and the concentration) in the samples  provided from the environment around the site. 

Abstract: make sure to include the following sections, where the background is about  

1. Background/problem => Aim 

2. Method 

3. Results 

4. Conclusions/outlook 

Introduction: 

Background to problem – can include things such as protein biotinylation, recombinant  expression, the accident etc.  

How problem may be solved – this is where you can bring in about use of ELISA to detect  presence and concentration of specific molecules (could perhaps compare with other  alternative techniques) 

Finish with Aim 

Results: 

Remember to write a narrative text section in Results that presents, describes and explains  briefly yor results – why you are doing each experiment, what was done, what was found  and briefly what it means and what it leads to next. Don’t just paste in graphs with no  explanation. 

With all figures, include a legend with heading and text explanation of the figure, not just a  key of symbols, such that figure can be understood without needing to refer to main text.  This legend should be added in Word document not in Excel but should be clearly attached  to the figure and easily distinguished from main text by font, size, italics etc. For example of  figures with legends, see Figure 1 below or attached published paper (Darwish et al 2020) including ELISA data in Figures 9, 10. Note, these legends could be improved as they don’t  explain the equation of the curves fitted to the data, the number of replicates in each data  point or what the error bars represent. 

Please don’t not include large data tables from Excel in your report. These should be  included as an appendix in the form of your Excel spreadsheet submitted as an attached file  with your report. 

Do not give numerical results with huge numbers of significant figure / decimal places.  Errors should be 1-2 sig figures only and main value with same decimal places eg. 13.6 ± 0.7,  not 13.5978543198 ± 0.71356420870. If there is no error given, use a reasonable number of  significant figure, usually 3-4 max. Note this applies to figure axes labels too – change to  round numbers if they come up by default as something else.

I think there is some confusion about Kd and Bmax because in the Equilibrium Dialysis  report both Kd and Bmax were measured as molar concentration (nM) and were in a similar  range. Kd should always be as a molar concentration but Bmax can be expressed in many  different units eg. ug/ml or simply as a % if normalised. The Kd is a property of the receptor  ligand interaction – ie the affinity - and is measured as a concentration that will give half the  maximal binding. The Bmax is simply dependent on the amount of receptor present. If you  increase the amount of receptor 10-fold, the Bmax will increase 10-fold but Kd will be  unchanged. For example, Figure 1. below. If you normalise binding on y-axis to a known  maximum as is done here, the Bmax becomes unitless, i.e. a fraction or %.

Some people have had concerns about 4PL fit parameters not being as expected. I had  mentioned that the Hill coefficient (Coeff b) is likely to be ~ 1 and negative for inhibition  curves. Don’t worry if it is quite different to 1. In MyCurveFit 4PL fitting to inhibition curves,  values for a and d are swapped – i.e. a is max and d is min - so Hill Coeff b stays positive. 

For point 10 in the Results, section of the Report Requirements, do not be concerned if your  calculated avidin binding greatly exceeds or is much less than the theoretical maximum.  

Finally, the marking scheme will follow the 100 marks in the online rubric (recently adjusted to  remove actual lab-work component) compared to the marks out of 40 listed in the prac manual but  proportions are roughly the same with an increased emphasis on results section.

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