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The Paragon CZE 2000 System identifies monoclonal components in serum by
immunofixation via subtraction (IFE/s). A patient serum sample is
automatically diluted and aliquoted into six wells of a specially
designed, sealed segment that contains buffer and antibody called
Sepharose®* beads. Five of the wells contain beads coated with antibody
specific to the heavy and light chain components of human
immunoglobulins (IgG, IgA, IgM, kappa, lambda). The sixth well, which
acts as a reference, contains buffer alone and produces a pattern
identical to the SPE profile. The serum sample is incubated with the
antibody-coated beads.
Immunoglobulins bind to the specific antibody coating the bead and are
subtracted out of solution as the bead settles to the bottom of the
well. For example, if IgA is present in a sample, the IgA will bind to
the Anti-IgA-coated bead and be pulled out of solution. When a component
is absorbed onto the beads it can be identified by the absence of the
corresponding peak in the electropherogram when compared to the SPE
reference pattern.
Comparisons of electropherograms will clearly show that IgA was present
in the sample, since its peak is absent when compared with that of the
reference.
The operator of the Paragon CZE 2000 System has the option of changing
the sample dilution prior to IFE/s analysis.
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