serum electrophoresis

8/7/2019 Serum Electrophoresis

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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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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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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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