Abstract
Immunoelectrophoresis (IEP) was the first practical method that combined electrophoresis and Âimmunoprecipitation for identifying and characterizing proteins within complex mixtures. Over the years, IEP has been extended to include a variety of techniques and, as a general name, has been applied to virtually any technique that involves electrophoresis and antigen–antibody precipitin reaction for proteins. Because of the diversity in technical details of different IEP versions, the method described here deals only with classic IEP. Although it requires some manual expertise, IEP is versatile, relatively easy to customize, and economical with no need for expensive instrumentation. Further, it can discern identity, partial identity, and nonidentity of the proteins. Any low-viscosity body fluid specimen or, possibly, culture fluid and tissue extract could be tested with IEP if proper antibodies are available. With these attributes, classic IEP remains a valuable tool for clinical diagnostic testing, purity checking of biochemical and pharmaceutical products, and research.
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References
Grabar P, Williams CA Jr (1953) Méthode permettant l’étude conjugée des propriétés électrophorétiques et immunochimiques d’un mélange de proteins. Application au serum sanguine. [Method permitting the combined study of the electrophoretic and the immunochemical properties of protein mixtures; application to blood serum]. Biochim Biophys Acta 10:193–194 [French]
Martin E, Scheidegger JJ, Grabar P et al (1954) [Immuno-electrophoretic study of blood serum]. Bull Schweiz Akad Med Wiss 10:193–198 [French]
Williams CA Jr, Grabar P (1955) [Immunoelectrophoretic method for the analysis of mixtures of antigenic substances]. Biochim Biophys Acta 17:67–74 [French]
Scheidegger JJ (1955) Une mict-méthode de l’immunoelectrophorése. Int Arch Allergy Appl Immunol 7:103–110 [French]
Scheidegger JJ, Roulet H (1955) [Practical application of the immuno-electrophoretic method; first results]. Praxis 44:73–76 [French]
Afonso E (1964) Quantitative immunoelectrophoresis of serum proteins. Clin Chim Acta 10:114–122
Wilson AT (1964) Direct immunoelectrophoresis. J Immunol 92:31–34
Bussard A (1959) [Description of a technic simultaneously combining electrophoresis and immunological precipitation in gel: electrosyneresis]. Biochim Biophys Acta 34:258–260 [French]
Vergani C (1971) Crossover electrophoresis for the rapid detection of serum hepatitis (Australia) antigen and antibody. J Clin Pathol 24:86–87
Tilton RC (1978) Counterimmunoelectrophoresis in biology and medicine. CRC Crit Rev Clin Lab Sci 9:347–365
Ressler N (1960) Two-dimensional electrophoresis of protein antigens with an antibody containing buffer. Clin Chim Acta 5:795–800
Laurell CB (1965) Antigen-antibody crossed electrophoresis. Anal Biochem 10:358–361
Clarke HG, Freeman T (1968) Quantitative immunoelectrophoresis of human serum proteins. Clin Sci 35:403–413
Laurell CB (1966) Quantitative estimation of proteins by electrophoresis in agarose gel containing antibodies. Anal Biochem 15:45–52
Laurell CB (1972) Electroimmuno assay. Scand J Clin Lab Invest Suppl 124:21–37
Weeke B (1969) [Quantitative immunoelectrophoresis. Rocket electrophoresis]. Ugeskr Laeger 131:1419–1423 [Danish]
Weeke B (1973) Rocket immunoelectrophoresis. Scand J Immunol Suppl 1:37–46
Svendsen PJ (1973) Fused rocket immunoelectrophoresis. Scand J Immunol Suppl 1:69–70
Bog-Hansen TC (1973) Crossed immuno-affinoelectrophoresis. An analytical method to predict the result of affinity chromatography. Anal Biochem 56:480–488
Kroll J (1973) Line immunoelectrophoresis. Scand J Immunol Suppl 1:61–67
Weeke B, Lowenstein H (1973) Allergens identified in crossed radioimmunoelectrophoresis. Scand J Immunol Suppl 1:149–153
Kroll J (1973) Crossed-line immunoelectrophoresis. Scand J Immunol Suppl 1:79–81
Kroll J (1973) Rocket-line immunoelectrophoresis. Scand J Immunol Suppl 1:83–87
Ohlenschläger G, Berger I (1974) [Tandem technic in Clarke, Freeman, Ressler and Laurell’s 2 dimensional immunoelectrophoresis. Methods and criteria for use as a consequence of the problems of realization of reproducible separations by stability of electrical values]. Z Med Labortech 15:119–126 [German]
Axelsen NH, Svendsen PJ (1973) Reversed rocket immunoelectrophoresis. Scand J Immunol Suppl 1:155–157
United States Pharmacopeial Convention (2011) Quality of biotechnological products: stability testing of biotechnological/biological products. In: The United States Pharmacopeia–National Formulary (USP 34–NF 29 through First Supplement). http://www.uspnf.com. Accessed 26 July 2011
European Directorate for the Quality of Medicines and Health Care (2012) Immunochemical methods. In: ep703 – European pharmacopoeia online, 7th Edition 2012 (7.3). http://online6.edqm.eu/ep703/. Accessed 26 July 2011
Nerenberg ST, Ganger C, DeMarco L (1972) Multi-antisera applicator for use in solid-support immunoelectrophoresis on cellulose acetate. Clin Chem 18:1021–1023
Arquembourg PC, Salvaggio JE, Bickers JN (1970) Primer of immunoelectrophoresis with interpretation of pathologic human serum patterns. Ann Arbor-Humphrey Science Publishers, Inc, Ann Arbor, MI [S. Karger, Basel]
Williams CA (1971) Immunoelectrophoretic analysis. In: Williams CA, Chase MW (eds) Methods in immunology and immunochemistry, vol 3. Academic, New York and London, pp 234–287
Cawley LP, Schneider D, Eberhardt L et al (1965) A simple semi-automated method of immunoelectrophoresis. Clin Chim Acta 12:105–110
Kostner G, Holasek A (1972) Influence of dextran and polyethylene glycol on sensitivity of two-dimensional immunoelectrophoresis and electroimmunodiffusion. Anal Biochem 46:680–683
Thoss K, Abendroth K (1974) [The effect of polyethylene glycol and dextran on the immunoelectrophoresis (author’s transl)]. Z Immunitatsforsch Exp Klin Immunol 147:404–412 [German]
Musset L, Diemert MC, Taibi F et al (1992) Characterization of cryoglobulins by immunoblotting. Clin Chem 38:798–802
Beyer NH, Schou C, Houen G et al (2008) Extraction and identification of electroimmunoprecipitated proteins from agarose gels. J Immunol Methods 330:24–33
Keren DF (1994) High-resolution electrophoresis and immunofixation. Techniques and interpretation, 2nd edn. Butterworth-Heinemann, Boston
Acknowledgments
The author wishes to thank Dr. Rita Ellerbrook of the Helena Laboratories, Beaumont, TX, USA for kindly providing illustrations to this article.
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Csako, G. (2012). Immunoelectrophoresis: A Method with Many Faces. In: Kurien, B., Scofield, R. (eds) Protein Electrophoresis. Methods in Molecular Biology, vol 869. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-821-4_28
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DOI: https://doi.org/10.1007/978-1-61779-821-4_28
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