Serum Protein Electrophoresis

Why Protein Electrophoresis

● Electrophoresis separates proteins based on
their physical properties, the subsets of these
proteins are used in interpreting the results.
● Used to identify patients with Multiple Myeloma
and other serum protein disorders.
● Commonly used to identify patients with
multliple myeloma and other serum protein
disorders.
● Sometimes results will be confusing to interpret.
 In this presentation
● How the examination is performed.

● What it measures and when it is indicated.

● Result interpretation

● Suggestion and follow-up of abnormal results

 Protein Electrophoresis
● Separtates proteins based on their physical
properties.
● Serum is placed in specific medium, and
charge is applied
● Based on the net charge & size and shape of
the proteins, proteins separate
● The pattern of serum protein electrophoresis
results depends on the fractions of two major
proteins: Albumin and Globulins.
 Protein Electrophoresis (Contd...)
● Several susbsets of serum protein can be separated
using different methods.
● In zone electrophoresis, different protein subtypes are
separated in different physical locations.
● Proteins are stained,and their densities are calculated
to provide graphical data on the absolute and relative
amounts of the various proteins.
● Further separation can be achived, by staining with
immunologically active agent, which results in
immunoflourescence and immunofixation.
 Components of Serum
Electrophoresis
● The pattern of serum protein electrophoresis
results depends on the fractions of two major
types of protein: Albumin and Globulins.
● Albumin – major protein component of serum,
is produced by the liver under normal
physiologic conditions.
● Globulins comprise of a much smaller fraction
of the total serum protein content.
● Subsets of globulins and their relative quantities
are the primary focus of interpretation of serum
protein electrophoresis.
 Normal pattern of Serum
Electrophoresis

+ ve Albumin
Globulins -ve
 Pattern of Serum Protein
Electrophoresis
● Closest to the Positive Electrode lies the largest
peak – Albumin.

● The next five components are Globulins –

Alpha1, alpha 2, beta1, beta 2 and gamma.

● The peaks lies towards negative electrode.

 Components: Albumin
● Represents the largest protein component of
human serum.
● Decreased albumin level is seen in case of
a. Less production of the protein by the liver.
b. Increased loss or degradation of this protein.
c. Dehydration.
● Malnutrition, significant liver disease, renal loss (e.g.
Nephrotic syndrome), hormone therapy, pregnancy
- may result in low albumin levels.
● Burns may result in increased levels of albumin.
 Components: Globulins
Alpha fraction of Globulins: Alpha 1, Alpha2
components.
Alpha 1 fraction is comprised of Alpha 1-
antitrypisin, thyroid- binding globulin, and
transcortin.
Malignancy and acute inflammation can
increase this band.
Decreased band may result because of
deficiency in Alpha 1- antitrypisin or liver disease
causing decreased level of globulin.
 Components: Globulins (contd...)
Second fraction of Alpha is the Alpha2 -protein.

● Ceruloplasmin, Alpha2-macroglobulin, and

haptoglobin are the components.

● Increased levels are seen as a result of Acute-

phase reaction.
 Components: Globulins (contd...)
Beta fraction of Globulin components have two
peaks Beta1 and Beta2.
● Beta1 is composed mostly of Transferrin.
● Beta2 contains Beta – lipoprotein.
IgA, IgM, and sometimes IgG, along
with complement proteins, also can be identified
in the beta fraction.
 Components: Globulins (contd...)
● Gamma Region of Electrophoretic region is of
great significance because of Immunoglobulin
migration in this region.
● Immunoglobulins often can be found throughout
the electrophoretic spectrum.
● C-reactive protein (CRP) is found located
between beta and gamma components.
 Interpretation of Results
Plasma protein levels display prodictable
changes in response to:
● acute inflammation,
● malignancy,
● trauma,
● necrosis,
● infraction,
● burns,
● chemical injury.
 Interpretation of Results (contd...)
● Interpretation mainly focus on the gamma
region, which is composed predominantly of
antibodies of IgG type.
● Gamma globulin zone is decreased in
hypogammaglobinemia and
agammaglobinemia.
● Diseases that produces an increase in the
gamma globulin level include Hodgkin's
disease, malignant lymphoma, chronic
lymphocytic leukemia, connective tissue
diseases, multiple myeloma, Waldenstrom's
macroglobulinemia, and amyloidosis.
 Abnormal Serum Protein Electrophoresis

● Several disease states cause an increase in

gamma region, due to several reasons a
homogeneous spike-like peak of the gamma
globulin zone. These may be due to
proliferation of a single clone of plasma cells
that produce a homogeneous M protein – called
“Monoclonal gammopathies”
 Monoclonal and Polyclonal
Gammopathies
● Monoclonal gammopathies are associated with a
clonal process that is malignant or potentially
malignant.
● Polyclonal gammopathies may be caused by any
reactive or inflammatory process and they are
associated with nonmalignant conditions.
● M-protein is charectarised by the prescence of a
sharp, well defined band with a single heavy chain and
a similar band with kappa and lambda light chain.
● A polyclonal gammopathy is charectarised by a broad
diffuse band with one or more heavy chains and kappa
and lambda light chains.