Immunoelectrophoresis

Immunoelectrophoresis is a general name for a number of biochemical methods for separation and characterization of proteins based on electrophoresis and reaction with antibodies.

[1] Agarose as 1% gel slabs of about 1 mm thickness buffered at high pH (around 8.6) is traditionally preferred for electrophoresis and the reaction with antibodies.

Among the important observations made were the great number of different proteins in serum, the existence of several immunoglobulin classes and their electrophoretic heterogeneity.

Affinity immunoelectrophoresis is based on changes in the electrophoretic pattern of proteins through specific interaction or complex formation with other macromolecules or ligands.

The open structure of the immunoprecipitate in the agarose gel will allow additional binding of radioactively labeled antibodies and other ligands to reveal specific proteins.

Today gel electrophoresis followed by electroblotting is the preferred method for protein characterization because its ease of operation, its high sensitivity, and its low requirement for specific antibodies.

In addition proteins are separated by gel electrophoresis on the basis of their apparent molecular weight, which is not accomplished by immunoelectrophoresis, but nevertheless immunoelectrophoretic methods are still useful when non-reducing conditions are needed.

The use of the fluorescein copolymer-antigen mixture improved the association with plasma levels antibodies of animals immunized against hemorrhage illness and enhanced protein concentration in the precipitated zone, according to the findings.

The capability of the amphiphilic fluorescein copolymer to boost antigen-antibody association and see the fluorescent accumulation domain may improve the efficiency of counter-immunoelectrophoresis for infectious disease rapid diagnosis.

[3] Immunomethods The terminologies, immune-methods and immune-chemical techniques refer to a variety of immunoelectrophoresis processes whose results are identified using antibodies and immunological methodologies.

Furthermore, in this procedure, the materials are placed into round wells in the gel's core part and disperse through it, generating a deposition ring with a diameter relation to the number of unbound protein that has diffused.

Traditional (classical or conventional) immunoelectrophoresis has a number of drawbacks, including the fact that it is time consuming and the protocol might take up to 3 days to finish, has limited specificity and sensitivity, and the results can be difficult to read.

Crossed immunoelectrophoresis of 2 microlitres of normal human serum. The electrophoresis was performed in thin layers of agarose; the pictured gel is about 7x7 cm. The lower part is the first dimension gel without antibodies, where the serum was applied into the slot at the lower left. The upper part is the second dimension gel with Dako antibodies against human serum proteins. More than 50 major serum proteins can be named.
Fused rocket immunoelectrophoresis of an affinity chromatographic separation of human serum proteins on con A . A 15 microlitre sample from each fraction was applied (starting from left) and allowed to diffuse about one hour, then electrophoresis was performed overnight. Peak "a" was not retained, while peak "b" was retained and eluted with methyl-mannose. The arrow indicates some slightly retained proteins.
Plasmodium Glutamate dehydrogenase (pGluDH) separated by Counterimmunoelectrophoresis [ 2 ]