Removal of the artifactual peak associated with concanavalin A crossed affinoimmunoelectrophoresis of human serum transferrin.
During the first dimension of crossed affinoimmunoelectrophoresis (CAIE) with the lectin concanavalin A (Con A), an immobile glycoprotein-Con A affinoprecipitate may form near the application well and, subsequently, produce an artifactual peak in the second-dimension gel. In this study, we examined the effects of sample glycoprotein concentration and gel Con A concentration on the magnitude of the transferrin artifactual peak present in the analysis of human serum. In addition, we examined the potential for reducing or eliminating the artifact by including a competitive inhibitor of glycoprotein-lectin interaction, alpha-methylmannoside (alpha MM), in the application well. We demonstrate that the artifact can be eliminated through an appropriate choice of glycoprotein, Con A, and alpha MM concentrations. This approach should be applicable for diagnosing and eliminating the artifact in the Con A CAIE analyses of other glycoproteins.
Hahn
T J
TJ
Department of Chemical Engineering, Stanford University, California 94305-5025.
Goochee
C F
CF
D016477:Artifacts; D003208:Concanavalin A / Q000378:metabolism*; D006023:Glycoproteins / Q000032:analysis*; D006801:Humans; D007122:Immunoelectrophoresis / Q000379:methods*; D008766:Methylmannosides / Q000378:metabolism; D014168:Transferrin / Q000032:analysis* / Q000276:immunology
D006023:Glycoproteins; D008766:Methylmannosides; D014168:Transferrin; D003208:Concanavalin A; C008466:methylmannoside