Immunoelectrophoresis

The Immunoelectrophoresis is used for laboratory diagnostic detection of monoclonal antibodies in a patient's blood. Monoclonal antibodies are derived from the same cell and are directed against the same antigens. For this reason, they are to be assessed as pathological and, if proven, speak for diseases such as Waldenström's disease.

What is Immunoelectrophoresis?

Ions have different mobility. This different mobility forms the basis of the electropheresis procedure. These methods separate different substances from one another using electric fields and gravity. A well-known method in the field is immunoelectrophheresis.

This is a qualitative diagnostic method that is used to detect monoclonal antibodies. Antibodies are immunologically active protein substances from a specific cell line. Monoclonal antibodies are all based on the same B lymphocytes and are therefore directed against a single epitope. Every natural immune response to invading antigens corresponds to a polyclonal response and is therefore directed against different epitopes. A monoclonal immune response therefore provides indications of pathological body processes.

Monoclonal antibodies bind different molecules with high specificity. This binding can be demonstrated by means of immunoelectrophoresis. The procedure is a qualitative procedure in laboratory diagnostics and consists of the two types of procedure, serum electrophoresis and immunodiffusion.

Function, effect & goals

Immunoelectrophoresis combines the methodology of serum electrophoresis with immunodiffusion. The patient's serum is placed on an agarose gel or cellulose acetate film. A control serum is also applied. The application is followed by an electrophoretic separation of the samples. Antisera, IgG, IgA, IgM, acetic acid for normal electrophoresis and kappa and lambda are applied between the dividing lines.

This creates a reaction with the antibodies in the patient's serum, which creates lines of precipitation. Depending on the antiserum used and the position and shape of the individual lines, conclusions can be drawn about the immunoglobulins contained with light chains kappa or lambda. In the case of a lambda band, there are free light chains made up of antibodies. The laboratory provides additional evidence via the rare IgE and IgD, which allows an exact determination of the immunoglobulins. A method of immunodiffusion electrophoresis takes place according to Pierre Grabar and Curtis Williams and corresponds to a combination of agarose gel electrophoresis of proteins and antibody diffusion.

First the agarose gel electrophoresis takes place. Subsequently, the contained antibodies diffuse against the bands of antigens and thereby create precipitate arches. This is to be distinguished from rocket immunoelectrophoresis according to Laurell, which corresponds to the electrophoresis of proteins within an agarose gel, each of which has antibodies in a certain concentration. There is a slightly basic buffer in the gel, which only allows the antigens to migrate and which forces most antibodies to the isoelectric point through exposure to the slightly basic pH value until they stop moving electrophoretically.

At the beginning of a rocket immunoelectrophoresis there is an excess of antigen, so that soluble antigen-antibody complexes are formed. During electrophoresis, there is also binding between antigens and other antibodies. In this way, immunoprecipitates are formed at the equivalence point, which resemble rocket-like figures with a height proportional to the antigen concentration. The height of the precipitate is measured to evaluate the test.

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Risks, side effects & dangers

The detection of monoclonal antibodies is of particular importance for the diagnosis of multiple myeloma and Waldenström's disease. The evidence provided speaks for a malignant degeneration of immune cells. Multiple myeloma corresponds to a cancer in the bone marrow, which is characterized by the malignant multiplication of antibody-producing cells in the plasma.

These plasma cells produce antibodies and their fragments. Malignant plasma cells always come from a common precursor cell and are therefore genetically identical. They only produce monoclonal antibodies. The malignancy of this disease can correspond to a precancerous stage, but it can also reach a highly malignant stage, which without treatment is quickly fatal. Symptoms of the disease result from the malignant growth of the cells or from the antibodies and antibody fragments. The most common symptoms include bone pain, bone dissolution, and spontaneous fractures.

The calcium level in the blood is often increased. In addition, the abnormal antibodies are often deposited in the tissue and cause functional disorders in the organs, which can lead to symptoms such as kidney failure or impaired blood flow. Waldström's disease is also a malignant tumor disease. More precisely, it is a malignant lymphoma disease that is counted among the slowly progressing and almost asymptomatic B-cell non-Hodgkin lymphomas. In most cases, abnormal production of monoclonal IgM due to the activity of the malignant lymphoma cells can be detected in the disease.

Waldenström's disease is extremely similar to multiple myeloma in its properties, but usually shows a more favorable course. Most patients with Waldenström's disease are largely asymptomatic by the time they are diagnosed. Other patients show symptoms early on, such as non-specific fatigue or peripheral neuropathies, which are caused by the deposition of monoclonal IgM within the myelin sheath. Even small amounts of IgM can trigger polyneuropathy. No polyneuropathy occurs in other patients, even at high levels. In addition, non-specific symptoms such as fever, unwanted weight loss or night sweats can occur. Bone pain is also characteristic.

Due to the overproduction of IgM, the blood becomes hyperviscous, so that the symptoms mentioned can be associated with those of a hyperviscosity syndrome. In most cases, this tendency to bleeding manifests itself in frequent nosebleeds, headaches, general malaise or blurred vision and acoustic complaints. In order to detect malignant diseases of this type, immunoelectrophheresis has long become a standard diagnostic procedure.