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Serum Protein Electrophoresis
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Why Protein Electrophoresis
Electrophoresis separates proteins based ontheir physical properties, the subsets of theseproteins are used in interpreting the results.
Used to identify patients with Multiple Myelomaand other serum protein disorders.
Commonly used to identify patients withmultliple myeloma and other serum protein
disorders.
Sometimes results will be confusing to interpret.
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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
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Protein Electrophoresis
Separtates proteins based on their physicalproperties.
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 electrophoresisresults depends on the fractions of two majorproteins: Albumin and Globulins.
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Protein Electrophoresis (Contd...)
Several susbsets of serum protein can be separatedusing different methods.
In zone electrophoresis, different protein subtypes areseparated in different physical locations.
Proteins are stained,and their densities are calculatedto provide graphical data on the absolute and relativeamounts of the various proteins.
Further separation can be achived, by staining withimmunologically active agent, which results inimmunoflourescence and immunofixation.
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Components of SerumElectrophoresis
The pattern of serum protein electrophoresisresults depends on the fractions of two majortypes of protein: Albumin and Globulins.
Albumin major protein component of serum,is produced by the liver under normalphysiologic conditions.
Globulins comprise of a much smaller fractionof the total serum protein content.
Subsets of globulins and their relative quantitiesare the primary focus of interpretation of serum
protein electrophoresis.
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Normal pattern of SerumElectrophoresis
+ ve Albumin
Globulins -ve
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Pattern of Serum ProteinElectrophoresis
Closest to the Positive Electrode lies the largestpeak Albumin.
The next five components are Globulins Alpha1, alpha 2, beta1, beta 2 and gamma.
The peaks lies towards negative electrode.
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Components: Albumin
Represents the largest protein component ofhuman 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.
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Components: Globulins
Alpha fraction of Globulins: Alpha1,Alpha2components.
Alpha1
fraction is comprised of Alpha1-
antitrypisin, thyroid- binding globulin, andtranscortin.
Malignancy and acute inflammation can
increase this band.Decreased band may result because of
deficiency in Alpha1- antitrypisin or liver disease
causing decreased level of globulin.
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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.
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Components: Globulins (contd...)
Beta fraction of Globulin components have twopeaks Beta
1and Beta
2.
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.
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Components: Globulins (contd...)
Gamma Region of Electrophoretic region is ofgreat significance because of Immunoglobulinmigration in this region.
Immunoglobulins often can be found throughoutthe electrophoretic spectrum.
C-reactive protein (CRP) is found locatedbetween beta and gamma components.
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Interpretation of Results
Plasma protein levels display prodictablechanges in response to:
acute inflammation,
malignancy, trauma,
necrosis,
infraction, burns,
chemical injury.
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Interpretation of Results (contd...)
Interpretation mainly focus on the gammaregion, which is composed predominantly ofantibodies of IgG type.
Gamma globulin zone is decreased in
hypogammaglobinemia andagammaglobinemia.
Diseases that produces an increase in the
gamma globulin level include Hodgkin'sdisease, malignant lymphoma, chroniclymphocytic leukemia, connective tissuediseases, multiple myeloma, Waldenstrom's
macroglobulinemia, and amyloidosis.
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Abnormal Serum Protein Electrophoresis
Several disease states cause an increase ingamma region, due to several reasons a
homogeneous spike-like peak of the gammaglobulin zone. These may be due toproliferation of a single clone of plasma cellsthat produce a homogeneous M protein called
Monoclonal gammopathies
M l l d P l l l
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Monoclonal and PolyclonalGammopathies
Monoclonal gammopathies are associated with aclonal process that is malignant or potentiallymalignant.
Polyclonal gammopathies may be caused by any
reactive or inflammatory process and they areassociated with nonmalignant conditions.
M-protein is charectarised by the prescence of asharp, well defined band with a single heavy chain anda similar band with kappa and lambda light chain.
A polyclonal gammopathy is charectarised by a broaddiffuse band with one or more heavy chains and kappa
and lambda light chains.
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