Abstract
An interaction or a binding event typically changes the electrophoretic properties of a molecule. Affinity electrophoresis methods detect changes in the electrophoretic pattern of molecules (mainly macromolecules) that occur as a result of biospecific interactions or complex formation. Lectin affinity electrophoresis is a very effective method for the detection and analysis of trace amounts of glycobiological substances. It is particularly useful for isolating and separating the glycoisomers of target molecules. Here, we describe a sensitive technique for the detection of glycoproteins separated by agarose gel-lectin affinity electrophoresis that uses antibody-affinity blotting. The technique is tested using α-fetoprotein with lectin (Lens culinaris agglutinin and Phaseolus vulgaris agglutinin)-agarose gels.
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References
Taketa K, Ichikawa E, Tago H et al (1985) Antibody-affinity blotting, a sensitive technique for the detection of α-fetoprotein separated by lectin affinity electrophoresis in agarose gels. Electrophoresis 6:492–497
Taketa K, Hirai H (1989) Lectin affinity electrophoresis of α-fetoprotein in cancer diagnosis. Electrophoresis 10:562–567
Kagebayashia C, Yamaguchi I, Akinaga A et al (2009) Automated immunoassay system for AFP-L3 % using on-chip electrokinetic reaction and separation by affinity electrophoresis. Anal Biochem 388:306–311
Shimura K (1990) Progress in affinophoresis. J Chromatogr 510:251–270
Shimura K, Kasai K (1995) Determination of the affinity constants of concanavalin A for monosaccharides by fluorescence affinity probe capillary electrophoresis. Anal Biochem 227:186–194
Shimura K, Kasai K (1996) Affinophoresis: selective electrophoretic separation of proteins using specific carriers. Methods Enzymol 271:203–218
El Rassi Z, Mechref Y (1996) Recent advances in capillary electrophoresis of carbohydrates. Electrophoresis 17:275–301
Kakehi K, Kinoshita M (2009) Capillary lectin-affinity electrophoresis for glycan analysis. Methods Mol Biol 534:93–105
Nakajima K, Kinoshita M, Oda Y et al (2004) Screening method of carbohydrate-binding proteins in biological sources by capillary affinity electrophoresis and its application to determination of Tulipa gesneriana agglutinin in tulip bulbs. Glycobiology 14:793–804
Nakajima K, Kinoshita M, Matsushita N et al (2006) Capillary affinity electrophoresis using lectins for the analysis of milk oligosaccharide structure and its application to bovine colostrum oligosaccharides. Anal Biochem 348:105–114
Taketa K, Fujii Y, Taga H (1993) Characterization of E-PHA-reactive alpha-fetoprotein isoforms by two-dimensional lectin affinity electrophoresis. Electrophoresis 12:1333–1337
Taketa K, Fujii Y, Aoi T et al (1993) Identification of alpha-fetoprotein-P4 sugar chain as alpha 2 → 6 monosialylated biantennary complex-type oligosaccharides with an exposed galactose on the mannose alpha 1 → 6 arm by two-dimensional extended agarose gel-lectin affinity electrophoresis. Electrophoresis 14:798–804
Taketa K, Ichikawa E, Sato J et al (1989) Two-dimensional lectin affinity electrophoresis of alpha-fetoprotein: characterization of erythroagglutinating phytohemagglutinin-dependent microheterogeneity forms. Electrophoresis 10:825–829
Taketa K, Sekiya C, Namiki M et al (1990) Lectin-reactive profiles of alpha-fetoprotein characterizing hepatocellular carcinoma and related conditions. Gastroenterology 99:508–518
Taketa K (1995) AFP. Rinsho Kensa 39:68–70
Yamashita K, Taketa K, Nishi S et al (1993) Sugar chains of human cord serum alpha-fetoprotein: characteristics of N-linked sugar chains of glycoproteins produced in human liver and hepatocellular carcinomas. Cancer Res 53:2970–2975
Kobayashi Y, Tateno H, Dohra H et al (2012) A novel core fucose-specific lectin from the mushroom Pholiota squarrosa. J Biol Chem 287:33973–33982
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Kobayashi, Y. (2014). Lectin Affinity Electrophoresis. In: Hirabayashi, J. (eds) Lectins. Methods in Molecular Biology, vol 1200. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1292-6_11
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DOI: https://doi.org/10.1007/978-1-4939-1292-6_11
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