Detection and quantification of ligand leakage from lectin affinity columns.
The usefulness of lectin affinity chromatography for the preparation of glycoproteins is impaired by ligand release. Ligand leakage from mistletoe lectin (MLI) Sepharose 4B column was detected by 24 h skin reaction in mice and by immunoblotting. Immunoaffinity chromatography was found to be an efficient method for the separation of lectin traces from the glycoprotein fraction. A sugar concentration dependent increase of lectin release from MLI-Sepharose 4B column was detected by a solid phase enzyme immunoassay.
Walzel
H
H
Institute of Biochemistry, Wilhelm-Pieck University, Rostock, GDR.
Brock
J
J
Franz
H
H
Ziska
P
P
D000818:Animals; D002846:Chromatography, Affinity; D005354:Fibrosarcoma / Q000032:analysis; D006023:Glycoproteins / Q000302:isolation & purification*; D037102:Lectins / Q000302:isolation & purification; D008024:Ligands; D051379:Mice; D008808:Mice, Inbred CBA; D014784:Mistletoe; D037121:Plant Lectins; D028321:Plant Preparations; D010940:Plant Proteins; D010946:Plants, Medicinal; D054790:Ribosome Inactivating Proteins, Type 2; D012513:Sarcoma, Experimental / Q000032:analysis; D012882:Skin Tests; D014118:Toxins, Biological / Q000302:isolation & purification / Q000633:toxicity
D006023:Glycoproteins; D037102:Lectins; D008024:Ligands; D037121:Plant Lectins; D028321:Plant Preparations; D010940:Plant Proteins; D054790:Ribosome Inactivating Proteins, Type 2; D014118:Toxins, Biological; C009674:ribosome inactivating protein, Viscum