Serum Protein Electrophoresis

What sample is needed for Serum Protein Electrophoresis?

1. This is done on the patient’s serum.
2. Also can be done in the urine. 24-hours of urine is preferred.
3. CSF for the oligoclonal band.

What are the indications for Serum Protein Electrophoresis?

1. This can diagnose some inflammatory diseases.
2. It can diagnose neoplastic diseases like multiple myeloma and lymphoma.
3. Advised In immune disorders.
4. Advised In liver diseases.
5. Advised In nephrotic syndrome.
6. Advised In Chronic edema.
7. To detect monoclonal proteins (Immunoglobulins).
8. CSF for the oligoclonal band.

How will you define electrophoresis?

1. This is the migration of charged solutes or particles in a liquid medium under the influence of the electrical field.
2. Analytes of interest are:
   1. Proteins.
   2. Peptides.
   3. Aminoacids.
   4. Nucleic acid.
   5. OLigonucleotides.
   6. Cations in body fluids and the tissue.

How will you explain the pathophysiology of serum proteins?

1. Proteins are constituents of muscles, hormones, hemoglobin, and transport proteins.
2. Serum proteins are a source of nutrition and a buffer system.
3. Immunoglobulins function in the immune system.
4. Carrier proteins, like haptoglobins, prealbumin, and transferrin, transport certain ions and molecules to their site of action.
5. Some of the protein regulates:
   1. The activity of various proteolytic enzymes.
   2. The osmotic pressure within the vascular compartment.
   3. Metabolic substances like a hormone.
6. Immunoglobulins are a major component of gamma-globulins.
   1. In a normal person, the immunoglobulins are polyclonal.
   2. The monoclonal band of immunoglobulin indicates a neoplastic process like multiple myeloma and Waldenstrom’s macroglobulinemia.
7. Serum electrophoresis separates proteins into 5 different bands:
   1. Albumin, and Prealbumin. Albumin is formed in the liver and is 60% of total proteins.
   2. alpha1-globulin (α₁-globulin).
   3. alpha 2-globulin (α₂-globulin).
   4. beta – globulin (β-globulin).
   5. gamma – globulin (γ-globulin).

What is the normal value of serum protein?

What are the patterns of different zones 0n Serum Protein Electrophoresis?

1. These different bands have different zones with the characteristic presence of different proteins. e.g.
   1. The Albumin zone shows only albumin.
   2. alpha1- zone shows:
      1. alpha1 – lipoprotein.
      2. High-density lipoprotein (HDL)
      3. Alpha-1 -antitrypsin.
   3. Alpha 2 zone shows:
      1. Alpha-2 macroglobulins.
      2. Haptoglobin.
      3. β -lipoprotein.
   4. Beta zone shows:
      1. Transferrin.
      2. Complement 3 (C3).
   5. Gamma zone shows
      1. Fibrinogen.
      2. IgA.
      3. IgM.
      4. IgG.

What are the functions of different proteins?

Albumin :

1. Maintain the colloidal osmotic pressure.
2. Albumin also transports drugs, hormones, and enzymes.
3. Albumin measures liver function. Its concentration is markedly low in liver diseases when liver cell damage occurs.
4. The half-life of albumin is 12 to 18 days so that liver cell damage will be observed after this period.

Globulins :

1. These are the main components of immunoglobulins (Antibodies).
2. Some are transporting proteins like thyroid and cortisol binding proteins.
3. Haptoglobin binds hemoglobin during hemolysis.
4. Ceruloplasmin is a carrier of copper.
5. They may also act as transport vehicles.

What is the Principle of Electrophoresis?

1. This is basically the separation or migration of charged solutes of particles in a liquid medium under the influence of the electrical field.
2. Chemical substances carrying charges because of ionization move toward either the cathode (Negative electrode) or the anode ( positive electrode). So, protein ions (cation) move toward the cathode, and negative ions (anion) move toward the anode.
3. Serum Electrophoresis can separate the various components of blood proteins into bands or zones according to their electrical charge under the influence of electrical current.
   1. Zones of proteins are separated from the neighboring zones.
   2. Then, these zones are visualized by the stains.
   3. The support medium is dried, and the densitometer can quantify each zone.
   4. The support medium can be kept permanently after drying.
4. The rate of migration depends upon the following:
   1. The net electrical charge of the molecule.
   2. Size and shape of the molecule.
   3. Electrical field strength.
   4. The temperature of the medium.
   5. Properties of the support medium.
5. Urine electrophoresis classifies renal protein-losing nephropathies.
6. CSF electrophoresis diagnoses the monoclonal band.

What components are needed for electrophoresis?

1. Tank with power supply (electrical field).
2. Electrode, either carbon or platinum.
3. Buffer system (in the buffer tanks).
4. Support media like :
   1. Agar gel.
   2. Agarose.
   3. Cellulose acetate.
   4. Polyacrylamide.
5. Stains. This is used to visualize the separate bands.
6. A densitometer is used to read the different bands and quantitate them.
7. The automated system automatically estimates electrophoresis bands instead of a manual method.

What are the normal Serum Protein Electrophoresis pattern?

Source 1

Total Proteins in Adult

• To convert into SI unit x 10 = g/L

Source 2

• Total protein = 6.4 to 8.3 g/dL.
• Albumin = 3.5 to 5 g/dL.
• Globulin = 2.3 to 3.4 g/dL.
  • alpha 1 globulin = 0.1 to .3 g/mL.
  • alpha 2 globulin = 0.6 to 1.0 g/dL.
  • beta globulin = 0.7 to 1.1 g/dL.

Children

• Total protein :
  • Premature infants = 4.2 to 7.6 g/dL.
  • Newborn = 4.6 to 7.4 g/dL.
  • Infants = 6.0 to 6.7 g/dL.
  • Child = 6.2 to 8.0 g/dL.
• Albumin :
  • Premature infants = 3.0 to 4.2 g/dL.
  • Newborn = 3.5 to 5.4 g/dL.
  • Infants = 4.4 to 5.4 g/dL.
  • Child = 4.0 to 5.9 g/dL.

Source 1

What is the pattern of Protein electrophoresis on cellulose acetate?

What is the pattern of serum Protein Electrophoresis on Agarose?

What are the causes of Increased Total Protein (Hyperproteinemia)?

1. Dehydration and hemoconcentration are due to fluid loss, such as vomiting, diarrhea, and poor kidney function.
2. Liver diseases.
3. Multiple myelomas.
4. Gammopathies.
5. Waldenstrom’s globulinemia.
6. Sarcoidosis and other granulomatous diseases.
7. Autoimmune diseases like SLE and Rheumatoid arthritis.
8. Chronic inflammation.

What are the causes of decreased Total protein (Hypoproteinemia)?

1. Severe liver disease.
2. Renal disease, nephrotic syndrome.
3. Starvation and malabsorption.
4. Diarrhea, ulcerative colitis, and Crohn’s disease.
5. Severe burn.
6. Skin diseases.
7. Hypothyroidism.
8. Heart failure.

What are the causes of increased Albumin?

1. Dehydration.
2. Intravenous infusion.

What are the causes of decreased Albumin?

1. Liver diseases.
2. Alcoholism.
3. Malabsorption.
4. Nephrotic syndrome.
5. Starvation.
6. Protein-losing enteropathies like Crohn’s disease and Whipple’s disease.
7. Ascites.
8. Congenital albuminemia.
9. Pregnancy.
10. Overhydration.
11. Autoimmune diseases like systemic lupus erythematosus.

What are the causes of increased α-1 globulin?

1. Biliary Cirrhosis.
2. Obstructive jaundice.
3. Multiple myelomas.
4. Kidney disease like nephrosis.
5. Acute and chronic infection.
6. Ulcerative colitis.

What are the causes of decreased α-1 globulin?

1. Kidney diseases like nephrosis.
2. Acute Hemolytic anemia.
3. Juvenile pulmonary emphysema.

What are the causes of increased α-2 globulin?

1. Kidney disease like nephrotic syndrome.
2. The inflammatory disease is due to an increase in the acute phase proteins.

What are the causes of decreased α-2 globulin?

1. In hemolysis, haptoglobin is alpha-2-globulin, which will decrease in hemolysis.
2. Wilson’s disease.
3. Hyperthyroidism.
4. Liver disease occurs when liver function is defective.

What are the causes of increased β- globulin?

1. Biliary Cirrhosis.
2. Obstructive jaundice.
3. Neoplasm-like multiple myeloma.
4. In all, hypercholesterolemia may be due to hypothyroidism and nephrosis.
5. Iron-deficiency anemia.

What are the causes of decreased β- globulin?

1. Kidney diseases like nephrosis.
2. Malnutrition.

What are the causes of Increased γ- globulin?

1. Hepatic diseases like cirrhosis.
2. Acute and chronic infections.
3. Multiple myelomas.
4. Autoimmune diseases like SLE and rheumatoid arthritis.
5. Waldenstrom’s macroglobulinemia.
6. Leukemia.
7. Malignancies like Hodgkin’s lymphoma and lymphomas.

What are the causes of decreased γ- globulin?

1. Nephrotic syndrome.
2. Agammaglobulinemia.
3. hypogammaglobulinemia.
4. An immune deficiency may be due to infections, steroid therapy, or lymphomas.

How will you explain Monoclonal Gammopathy?

• It is seen in some malignant processes like multiple myeloma.

• Please see more details on Protein Total.