Immunofixation electrophoresis

Capillary electrophoresis subsequent to immunosubtraction, which entails incubating the serum sample with beads coated with specific antisera followed by analysis of supernatants, is an attractive alternative to immunofixation and is amenable to automation (K36). 

D. F. Keren, High Resolution Electrophoresis and Immunofixation, Butterworths, London, 1987. 

Keren DF. High-resolution electrophoresis and immunofix-ation. Techniques and interpretation. Oxford Butter-worth-Heinemann, 1994. 

In clinical laboratory medicine, this procedure has been applied to the evaluation of human myeloma proteins. However, the method is being replaced by immunofixation (IF) electrophoresis, particularly for the study of protein antigens and their spHt products, and for evaluation of human myeloma proteins (see later discussion of IF electrophoresis). 

Alper CA, Johnson AM. Immunofixation electrophoresis A technique for the study of protein polymorphism. Vox Sang 1969 17 445-52. 

Keren DE High Resolution Electrophoresis and Immunofixation Techniques. Boston Butterworth-Heineman, 1994. 

There are several genetic variants or isotypes of Cp, but none of these, except for genetic deficiency (see previous discussion in this chapter), has known clinical significance. The most prevalent isotypes are CpA, CpB, and CpC, which are detectable by electrophoresis followed by either immunofix-ation or functional staining CpB is the common, or "wild, isotype. The gene encoding Cp is on chromosome 3q25. ... 

Electrophoresis is used to study and measure the protein content of biological fluids (see Chapter 5). Types include serum protein electrophoresis using either cellulose acetate strip or agarose gel as the separation media, capillary electrophoresis (CE), immunofixation, and Western blotting. ... 

Immunofixation electrophoresis (IFE) is gradually replacing immunoelectrophoresis (lEP) for detection of. M-components because of its speed and ease of interpretation. Several procedures, including commercial Idts, are available for both IFE and lEP, Although these procedures may differ in detail, their principles are similar. 

Figure 20-10 Comparison of immunofixation electrophoresis (IFE) and immunoelectrophoresis (lEP) for two patients with monoclonal gammopathies. A, Patient specimen with an IgG (kappa, k) monoclonal protein as identified by IFE.The arrow indicates the position of monoclonal protein.After electrophoresis, each track except SPE is reacted with its respective antiserum, then all tracks are stained to visualize the respective protein bands. (SPE Chemically fixed serum protein electrophoresis IgG, IgA, IgM, k, and A, indicate antiserum used on each track.) B, Same specimen as in A, with proteins identified by lERThe arrow indicates the position of monoclonal protein. Norma control (C) and patient sera (S) are alternated. After electrophoresis, antiserum is added to each trough as indicated by the labels ig (polyvalent Ig antiserum), IgG, IgA, IgM, K, and A The antisera react with separated proteins in the specimens to form precipitates in the shape of arcs.The IgG and k arcs are shorter and thicker than those in the normal control, showing the presence of the IgG (k) monoclonal protein.The concentrations of IgA, IgM, and A,-iight chains are also reduced. C, Patient specimen with an IgA (lambda, A,) monoclonal protein identified by IFE procedure as described in A. D, Same specimen as in C with proteins identified by lEP as described in B.The abnormal IgA and A,-arcs for the patient specimen indicate an elevated concentration of a monoclonal IgA (A.) protein. Ait separations were performed using the Beckman-Coulter Paragon system.

Serum immunofixation electrophoresis of two patients with monoclonal gammopathies. Pattern A represents IgG(/c) monoclonal gammopathy and pattern B represents IgA(/r) monoclonal gammopathy, as indicated by arrows. The procedure consists of serum protein electrophoresis (SPE) separation, reaction of each track with the exception of SPE with specific respective antiserum, followed by protein staining to make visible the respective bands. 

Levinson SS. Urine protein electrophoresis and immunofixation electrophoresis supplement one another in characterizing proteinuria. Ann Clin Lab Sci 2000 30 79-84. 

Harrison, H.H., Miller, K.L., Abu-Alfa, A., and Podlasek, S.J., 1993, Immunoglobulin clonality analysis Resolution of ambiguities in immunofixation electrophoresis results by high-resolution, two- dimensional electrophoretic analysis of paraprotein bands eluted from agarose gels. m. J. Clin. Pathol. 100 550-560. 

Figure 1 Properdin factor-B phenotyping performed using thin-layer high-voltage agarose gel electrophoresis and subsequent immunofixation with specific antiserum.

Techniques for the detection or assay of various substances based on the reaction of those substances with specific antibodies (or vice versa, i.e. the detection and assay of antibodies using antigens). Such techniques include agglutination reactions, automated immune precipitation, complement fixation tests, crossed electrophoresis, counter electrophoresis, double diffusion, enzyme immunoassay, fluoroimmunoassay, haem-agglutination, immunoelectrophoresis, immunofluoresence, radial immunodiffusion, spin immunoassay, immunofixation, immunoradiometric assay and radioimmunoassay. See separate entries for these subjects. 

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