antibody titration analysis

Eric Martz emartz at microbio.umass.edu
Wed Nov 6 08:37:15 EST 1996


On 11/5/96 Tom McHugh wrote:
>
>Are there any recommendations of available PC programs for the analysis
>of antibody titration experiments? I am using flow cytometry to detect
>specific antibody in serial dilutions of serum containing polyclonal antibody
>to three separate antigens. I am interested in comparing "titer" among
>three groups: (1) antibody negative (non-immunized), (2) strong antibody
>positives, (3) negative or weak antibody (immunized but immunosuppressed).

Dear Tom:  MFI (free DOS program for PC's) was written to do exactly this,
processing large numbers of listmode files in batch mode quickly, getting
corrected intensities.  It comes with a tutorial and sample titration files
which illustrate this use.  For monoclonal antibodies, the results usually
fit the one site binding equation nicely, I/Imax = C/(C + Ch), where I is
intensity, C is concentration (e.g. v/v %), and Ch is the concentration
which half-saturates.  MFI doesn't do this curve fit, but it can put its
results into a spreadsheet ready file, and the curve fit can be done in any
generic scientific data plotting program.  Polyclonal antibodies, not
surprisingly, usually don't fit this equation well.  Check out MFI at the
free software catalog, URL below.
/* - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
Eric Martz, Professor (Immunology)
Dept Microbiology, Univ Massachusetts, Amherst MA US 01003-5720
413-545-2325 FAX:545-1578.  Web projects include:
RasMol Home Page, http://www.umass.edu/microbio/rasmol
Catalog of Free Flow Cytometry Software (includes MFI which I wrote),
  http://www.bio.umass.edu/mcbfacs/flowcat.html
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