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Protein Electrophoresis Serum

Serum proteins have different net charges and can be separated by electrophoresis into several distinct bands. The band that migrates fastest toward the anode is albumin followed by alpha 1-globulin, alpha 2-globulin, beta globulin, and gamma globulins.  Protein concentrations may be altered as a result of different disease states.  Interpretation of serum protein electrophoretic patterns is helpful in diagnosing some diseases The most commonly recognized electrophoretic patterns are acute inflammation, alpha-1 antitrypsin deficiency, chronic inflammation, cirrhosis, hypoalbuminemia, hypogammaglobulinemia, monoclonal gammopathy, polyclonal gammopathy and protein losing disorder.

 

Pattern

Protein Changes

Frequently Associated Diseases

Acute

Inflammation

Normal or Decreased albumin

Increased a1G &/or a2Globulin

Acute infection and inflammatory disorders

Chronic

Inflammation

Normal or Decreased albumin

Increased a1G or a2Globulin

Increased gamma Globulin

Autoimmune diseases, chronic liver disease, chronic infection, cancer

Hypo-

albuminemia

Decreased albumin

Metastatic cancer, CHF, malnutrition, protein losing disorders

Hypogamma-globulinemia

Normal or decreased albumin

Decreased gamma Globulin

Lymphoproliferative disorders, inflammatory bowel disease, congenital immunodeficiencies

Polyclonal gammopathy

Increased gamma Globulin

Autoimmune disease, infections,

Liver disease

Cirrhosis

Increased gamma Globulin

Beta-gamma bridging

Cirrhosis

Protein losing disorder

Decreased albumin

Decreased alpha 1Globulin

Increased alpha 2Globulin

Increased beta Globulin

Nephrotic syndrome,  exudative skin disorders,  gastroenterophathies,

 

Monoclonal gammopathy

Normal or Decreased albumin

Increased gamma Globulin

Myeloma, macroglobulinemia, MGUS, CLL, lymphoma

Antitrypsin deficiency

absent alpha 1Globulin

Alpha 1 antitrypsin deficiency

Hyperbeta-

globulinemia

Normal – Decreased albumin

Increased beta Globulin

Hyperlipidemia, diabetes mellitus,

iron deficiency anemia

 

SPE Interpretation Tips

  • Albumin band often looks less intense on IFE compare to SPE, probably because it is incompletely fixed and elutes off the gel
  • Bisalbuminemia is most commonly seen in Native American individuals
  • Bilirubin, heparin & antibiotic binding can cause slurring of albumin band
  • Hemolysis causes decreased alpha-2 band (haptoglobin) and appearance of Hb band between alpha-2 and beta-1 regions
  • C3 is labile & decreases with storage; results in much variation in beta-2 region
  • Fibrinogen migrates between beta-2 and gamma regions (close to application point) and is present in plasma or heparin contaminated specimens
  • Immune complexes appear as monoclonal band at application point
  • Monoclonal bands in the beta region may indicate light chain disease, amyloidosis, heavy chain disease, IgD and IgE monoclonal gammopathies
  • Gamma heavy chain disease can produce a relatively broad band anywhere from the alpha-2 through the gamma region
  • CRP migrates in the gamma region and resembles a monoclonal band; present in acute inflammation
  • High resolution SPE gel can detect a monoclonal band of 0.1 g/dL
  • High resolution SPE cannot accurately quantitate monoclonal bands <0.3 g/dL
  • Oligoclonal bands with hypergammaglobulinemia & possibly beta-gamma bridging may be present in serum in patients responding to antigenic stimulation resulting from viral & bacterial infections, vaccines, autoimmune diseases and angioimmunoblastic lymphadenopathy. 
  • Oligoclonal bands with decreased IgG level is seen in CLL, post heart and BM transplants, and common variable immunodeficiency and immunosuppressive Rx.
  • Infections may cause transient monoclonal proteins
  • Light chain disease can result in monoclonal kappa or lambda chains in serum and not in urine if light chains are polymerized
  • Kappa chains usually stain more strongly than lambda chains
  • Broader width of monoclonal bands may be related to amount of protein applied to gel or heterogeneity of monoclonal protein due to glycosylation. IgA monoclonal bands are usually broader than IgG.
  • A monoclonal band may be clinically significant if it is at least as intense as the alpha 1 band.

Clues that a monoclonal band is unlikely to be due to a malignant clonal expansion

  • Acute phase pattern is present along with monoclonal band
  • Monoclonal band is transient & may evolve into an oligoclonal pattern
  • All immunoglobulin classes are elevated along with monoclonal
  • Slightly abnormal kappa:lambda ratio
  • Bence Jones protein is absent from urine

 Serum protein electrophoresis should be repeated in one year for asymptomatic patients with a monoclonal protein less than 1.5 g/dL and normal values of hemoglobin, calcium, and creatinine.  Electrophoresis should be repeated in two to three months if the monoclonal protein is between 1.5 and 2.5 g/dL.  Patients being treated for multiple myeloma, Waldenstrom’s macroglobulinemia or amyloidosis should be monitored at one to two month intervals. 

 Reference ranges using the SPIFE 4000 Split Beta SPE system are:

 

Protein Fraction

Reference Range

Total Protein

6.0 – 8.0 g/dL

Albumin

3.4 – 5.0 g/dL

Alpha 1 globulin

0.2 – 0.4 g/dL

Alpha 2 globulin

0.5 – 1.1 g/dL

Beta globulin

0.7 – 1.5 g/dL

Gamma globulin

0.5 – 1.5 g/dL

 

Specimen requirement is a red top tube of blood.

 

 

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