Protein Electrophoresis and Immunofixation Electrophoresis
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A protein electrophoresis and immunofixation electrophoresis test measures and separates different proteins in the blood, urine or sometimes other body fluids using samples such as a blood sample taken from a vein in the arm. It is used to help detect, diagnose and monitor conditions associated with abnormal protein production, including multiple myeloma and other disorders affecting the immune system, kidneys or liver.
Why get tested?
To help diagnose and monitor tumours of the antibody producing cells (B lymphocytes) and to diagnose and monitor deficiencies of the immune system
When to get tested?
If your doctor suspects that you have a condition that affects the antibody concentrations in the blood. If you have no symptoms is may be that other test results suggest a problem with antibodies such as a high total protein concentration
Sample required?
A blood sample taken from a vein in your arm; sometimes a random or 24-hour urine sample
Test preparation needed?
No test preparation is needed
What is being tested?
Protein electrophoresis is a method for separating the proteins found in blood (serum) or urine. During the test, an electric current is used to move the proteins across a thin layer of agarose gel. The distances that individual proteins travel depend on their size, shape, and electrical charge. These separated proteins may be detected by the use of a dye that binds to (stains) all of the proteins and reveals a characteristic pattern of bands. Each band indicates the presence of a particular protein or group of proteins, while the size of the band is a rough indication of the quantity. This pattern of bands is converted into a visual graph, showing vertical spikes or peaks where there is a lot of protein and smaller peaks or valleys where there is less. An alternative method called capillary zone electrophoresis (CZE) separates proteins by passing them through a long, thin column, producing a graph that is very similar to the one made by running the protein through an agarose gel.
Specific proteins of interest can be identified by separating the proteins by gel electrophoresis and then adding an antibody. The antibody can recognise individual proteins and sticks to them making a big complex that gets caught in the gel. All the other proteins in the system are washed away and the gel stained so the proteins in question can be identified. This procedure is called immunofixation electrophoresis (IFE) and only performed if the original electrophoresis is abnormal.
The bands seen on protein electrophoresis form characteristic patterns. Alterations to these patterns are associated with a variety of different diseases and conditions. For example in multiple myeloma (a cancer of certain types of white blood cells called plasma cells), the uncontrolled growth and division of a malignant plasma cell leads to the production of large amounts of a single type of immunoglobulin (antibody). In contrast to other proteins in serum, which are typically of a single type, antibodies (immunoglobulins) must differ from each other to be able to recognise bacteria, viruses and other ‘foreign’ substances. Each time the body is exposed to a virus, for example, one plasma cell replicates and makes a group (or clone) of plasma cells to produce antibody to eliminate it. Since our total immunoglobulin represents antibody made by many clones, we refer to it as a polyclonal pattern. When there is a cancer of plasma cells, only one type of antibody is produced, termed a monoclonal pattern. This abnormal protein can be seen as a characteristic band on the electrophoresis gel.
Common questions
Electrophoresis is used to identify the presence or absence of abnormal proteins and to identify when different groups of proteins are increased or decreased in serum or urine. It is frequently used to detect and identify monoclonal proteins (an excessive production of one specific immunoglobulin). Protein and immunofixation electrophoresis are used to help detect, diagnose, and monitor the course and treatment of conditions associated with these abnormal proteins, including multiple myeloma and a few related diseases.
Protein is usually excreted in the urine in very small amounts. When it is present in moderate to large amounts, it often indicates a problem with the kidneys. The primary reason protein and immunofixation electrophoresis are requested on urine is to look for monoclonal protein production which has largely been replaced by serum free light chain analysis.
Protein electrophoresis may be requested when a doctor is investigating symptoms that suggest multiple myeloma, such as bone pain, anaemia, tiredness, unexplained fractures, and recurrent infections. It may also be used as a follow-up to other laboratory tests, such as an abnormal total protein and/or albumin level, elevated urine protein levels, elevated calcium levels, and low white or red blood cell counts. Immunofixation electrophoresis is usually ordered when the protein electrophoresis test shows the presence of an abnormal protein band that may be an immunoglobulin.
Electrophoresis tests are most frequently requested when a doctor suspects a disease or condition that causes a monoclonal protein to be produced. Once a disease or condition has been diagnosed, electrophoresis may be used at regular intervals to monitor the course of the disease and the effectiveness of treatment. As disease progresses, the amount of protein goes up; with treatment (if required), it goes down. Monoclonal protein production may be due to a malignant disease, such as multiple myeloma, but it may also be due to a monoclonal gammopathy of undetermined significance (MGUS). Most patients with MGUS have no symptoms, but they must continue to be monitored regularly as some may develop multiple myeloma after a number of years.
Serum protein electrophoresis may also be used when symptoms suggest an inflammatory condition, an autoimmune disease, an acute or chronic infection, a kidney or liver disorder, or a protein-losing condition, even if the total protein and/or albumin concentrations are apparently normal.
The main use of electrophoresis is for detection of possible myeloma, therefore although many other protein patterns can be seen the reporting is usually limited to whether a monoclonal band is identified or not and what type of antibody it is and how much.
Immunisations within the previous six months can increase immunoglobulins as can drugs such as phenytoin (Dilantin), procainamide, oral contraceptives, methadone, and therapeutic gamma globulin.
Aspirin, bicarbonates, chlorpromazine (Thorazine), corticosteroids, neomycin and drugs based on antibodies can affect protein electrophoresis results.
They may do a serum electrophoresis because you may have an abnormality even though the total protein and albumin are normal. This is because the body tries to maintain a constant amount of protein and may increase or decrease its production of other proteins to compensate for a deficiency or overproduction of others.
Yes, any time separation of molecules is desired. DNA electrophoresis, for instance, is used to help study the genetic makeup of plants, animals, and humans.