Immunoelectrophoresis

Immunoelectrophoresis is a qualitative method for the detection of monoclonal antibodies in laboratory medicine.

Principle

Immunoelectrophoresis ( immunoelectrophoresis ) combines two methods with each other: the serum electrophoresis and immunodiffusion. On an agarose gel, on a cellulose acetate film rare patient serum (or the to be assayed sample of antigens ) is first applied and a control serum and separated by electrophoresis. Subsequently, between the two dividing lines antisera acetic acid applied for the normal electrophoresis, IgG, IgM, IgA, kappa or lambda. This reacts with the antibodies in the patient serum or control serum and forms characteristic precipitin. Depending on the nature of the antiserum, the position and shape of the lines may be used to infer the presence of the immunoglobulin kappa and lambda light chains. If in the example described only a lambda band is seen, either free light chains of antibodies may be present in the rarer IgE and IgD, another evidence must be provided in order to determine to which of the two immunoglobulins it is. In biochemistry and cell biology further, the investigated substances adapted variants are in use.

Special methods

Immunodiffusion electrophoresis according to Grabar and Williams

The immunodiffusion electrophoresis by Pierre Grabar and Curtis Williams is a combination between agarose gel electrophoresis of the proteins ( antigens) and a diffusion of their antibody dar. After agarose gel electrophoresis diffuse the antibodies, which are inserted into punched gutters, against the antigen bands and form with them Präzipitatbögen ( comparable to the precipitation lines ).

Rocket immunoelectrophoresis according to Laurell

The rocket immunoelectrophoresis by Carl- Bertil Laurell ( Lund University ) is an electrophoresis of proteins (antigen) in an agarose gel containing antibodies to a given concentration. The buffer in the gel is slightly basic, so that only the antigens can migrate because most antibodies are at a slightly basic pH at their isoelectric point and therefore can not move electrophoretically. At the beginning of rocket immunoelectrophoresis exists an antigen excess, leading to formation of soluble antigen-antibody complexes. During electrophoresis, the antigens bind additional antibodies and form at the equivalence point immunoprecipitates. These resemble rocket shaped figures, the surface (height) is proportional to the antigen concentration. For evaluation, one misses only the height of the precipitate.

Indication

Detection of monoclonal antibodies by immunoelectrophoresis is of particular importance in the diagnosis of multiple myeloma or Waldenstrom 's disease, but also in other diseases with malignant degeneration of immune cells.

410354
de