spiking antibody into sera slide 03-24-2014d1544

slide
-"C:\Users\kurtw_000\Documents\kurt\storage\CIM Research Folder\DR\2014\03-24-2014d0913\Spiking antibody into sera 2.pptx"
-"C:\Users\kurtw_000\Documents\kurt\storage\CIM Research Folder\DR\2014\03-24-2014d0913\Spiking antibody into sera.pptx"

some powerpoints this slide is found in
-C:\Users\kurtw_000\Documents\kurt\storage\CIM Research Folder\kwhittem\Presentations\2014\PhD Oral Defense\Oral Defense Presentation Kurt Whittemore 4-11-14.pptx

text from slide
{
Summary Points
-experiment by Josh Richer
-polyclonal mouse sera against the human GFOD1 protein
-two technical replicates
-CIM10kv2 arrays
-complexity of sera may change with an acute infection or during a B cell lymphoma
-decrease in antibody repertoire complexity should result in a decrease in fluorescence intensity distribution complexity
-surprise with possible saturation and “spill over”
-another experiment from Heidi and Bart support these results (not shown)

Some text from dissertation related to this experiment
{
In the previous example, two antibodies were applied to the array separately and in a mixture, but this is far from a practical case.  In a real situation, the blood will contain a high diversity of many different antibodies since there are a total of about 109 different antibodies in sera.  Additionally, the complexity of a whole antibody repertoire can change as a few single antibodies against a few targets may come to dominate the mixture.  This scenario can occur when there is a strong immune response against a single virus or bacteria type.  Alternatively this scenario could also occur when a lymphoma causes a single B cell to proliferate out of control and produce many antibodies against a single target.  In these scenarios, the complexity of the antibody repertoire is reduced as copies of antibodies against a single target make up an increasingly greater portion of the antibody repertoire.  This type of situation was simulated by spiking increasing concentrations of monoclonal antibody into sera.  The expectation is that a reduction in the antibody repertoire complexity that interacts with the peptide microarray will result in a reduction in complexity in the fluorescence intensity distribution.  A lower complexity fluorescence intensity distribution will result in changes in the AbStat measures.  For example, the entropy will decrease as the concentration of monoclonal antibody in the sera is increased.
Two experiments were performed in which antibody against a particular target was added to normal sera in increasing concentrations.  In the first experiment, polyclonal mouse sera against the human GFOD1 protein was added to normal mouse sera.  In the second experiment, human monoclonal antibody against the human gp120 HIV protein was added to normal human sera.
Polyclonal sera against the human GFOD1 (glucose fructose oxidase 1) protein was added to normal mouse sera at increasing concentrations by Josh Richer.  The polyclonal IgG mouse antibody was added to a 1:500 dilution of normal mouse sera in order to obtain the following final antibody concentrations: 0.1 nM, 1 nM, 2.5 nM, 5 nM, 10 nM, and 40 nM.  There were two technical replicates for each condition applied to the CIM10Kv2 arrays.  A line graph of the entropy vs antibody concentration (Figure 9) and a line graph of the mean vs antibody concentration (Figure 10) actually reveals that the two graphs follow a similar pattern which decreases up to 5 nM and then starts increasing.  The order of the measures with the most significant p-values between normal mouse sera and 5 nM antibody is as follows: 95th percentile, 5th percentile, median, mean, entropy, and cv.  The concentration of 5 nM was chosen as the comparison point because this is the point at which the trend in the curves reverses.
Figure 9 Entropy for increasing antibody concentrations in normal mouse sera
Figure 10 Mean for increasing concentrations of normal mouse sera

}

}