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  • Biochemistry and Biotechnology  (2)
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  • 1
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 10 (1989), S. 841-847 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: In contrast to the conventional combination of physical, chemical and enzymatic methods used for a structural analysis of glycans in glycoproteins, alternative methods involve affinity electrophoresis as a tool for the detection, characterization, and quantitation of glycoproteins and their carbohydrate moiety, owing to interactions with lectins. Two major approaches involve (i) crossed affino-immunoelectrophoresis and variations thereof, whereby lectin/glycoprotein interactions occur during the electrophoretic runs, or (ii) affino-blotting, where the glycoproteins are electrophoretically separated and then immobilized onto a solid support prior to their interaction with lectins. A critical comparison of these two series of techniques is the scope of the present paper. These techniques are of high interest by virtue of their ability at differentiating a classical glycan structure from unusual oligosaccharide side chains. The former structures will usually be qualitatively and quantitatively described with the easy and fast procedures as well as the simple equipment required for crossed affino-immunoelectrophoresis or affino-blotting, whereas the latter will be good candidates for further structural analyses.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Crossed affinoimmunoelectrophoresis (CAIE) with free lectin in the first dimension was used to investigate sugar heterogeneity of inter-α-trypsin-inhibitor (ITI) in normal human serum and after purification. Such a heterogeneity was assessed with Lens culinaris agglutinin and Concanavalin-A, but not with wheat germ agglutinin. The use of a mixture of purified ITI and ITI-free serum demonstrated that a distortion of the immunoprecipitation patterns for ITI in normal serum was observed with Con-A and WGA and was due to presence of other glycoproteins. Thus, prior protein purification is sometimes required before obtaining a true pattern of glycoprotein microheterogeneity by lectin CAIE. Furthermore, loading a sugar specific for the lectin in the second dimension gel was shown to be necessary in order to observe all heterogeneous forms of the glycoprotein tested.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
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