Electrophoresis Techniques

Immunoelectrophoresis
Immunofixation Electrophoresis
Electrophoresis Techniques
(Serum Electrophoresis)
Serum electrophoresis
results in the separation
of proteins into five
fractions on cellulose
acetate.


Electrophoreisis Techniques
(Immunoelectrophoresis)
By comparison to
serum electrophoresis,
immunoelectrophoresis
involves the
electrophoresis of
serum or urine
followed by
immunodiffusion.

Electrophoresis Techniques
(Immunoelectrophoresis)
IEP is a reliable and accurate
method for detecting both structural
abnormalities and concentration
changes in proteins.
The most common application of
IEP is in the diagnosis of a
monoclonal gammopathy, a
condition in which a single clone of
plasma cells produces elevated
levels of a single class and type of
Ig.
The most important application of
IEP of urine is the demonstration of
Bence Jones Protein. (kappa or
lambda light chains)

Electrophoresis Techniques
(Immunoelectrophoresis)
The size and position of
precipitin bands provide
the same type of
information regarding
equivalence or Ag-Ab
excess as double
immunodiffusion systems.
The position and shape of
precipitin bands in the IEP
assay of serum are
relatively stable and
reproducible. Any
deviation is abnormal.
These abnormalities can be
detected by evaluating the
following features of the
precipitin bands:
Position of the band
Distortion of the curvature
or arc formation
Thickening (density) and
elongation of a band
Shortening (inhibition),
thinning, or doubling.

Electrophoresis Techniques
(Immunofixation electrophoresis)
Immunofixation
electrophoresis (IFE) is
a two-stage procedure
using agarose gel
protein electrophoresis
in the first stage and
immunoprecipitation in
the second.


Electrophoresis Techniques
(Immunofixation electrophoresis)
The tests specimen
may be serum, urine,
CSF, or other body
fluids
The primary use of IFE
in clinical laboratories
is for the
characterization of
monoclonal Igs.

Comparison of IEP and IFE
IEP is technically simpler
and less subject to Ag
excess than IFE.
IFE high concentrations of
monoclonal protein give no
visible reactions, IEP is
considered to be a better
technique for typing large
monoclonal gammopathies.
IFE can be optimized to
give both greater
sensitivity and resolution
than IEP.
IFE should be reserved for
anomalous proteins, which
are difficult to characterize
by IEP.
IFE is easier to interpret
than IEP.