Analysis of the fine specificity of Tn-binding proteins using synthetic glycopeptide epitopes and a biosensor based on surface plasmon resonance spectroscopy

• Published in: FEBS letters Vol. 469; no. 1; pp. 24 - 28
• Main Authors: Osinaga, Eduardo, Bay, Sylvie, Tello, Diana, Babino, Alvaro, Pritsch, Otto, Assemat, Karine, Cantacuzene, Daniele, Nakada, Hiroshi, Alzari, Pedro
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 03.03.2000; Wiley
• Subjects: Animals; Antibodies, Monoclonal - immunology; Antigens, Tumor-Associated, Carbohydrate - chemistry; Antigens, Tumor-Associated, Carbohydrate - immunology; Biomarkers, Tumor - immunology; Biosensing Techniques; Cancer; Epitopes - chemistry; Glycopeptides - chemical synthesis; Glycopeptides - immunology; Glycosylation; Kinetics; Lectin; Lectins - immunology; Life Sciences; mAb, monoclonal antibody; Mice; Monoclonal antibody; O-Glycosylation; Plant Lectins; Protein Binding; Surface Plasmon Resonance; Surface plasmon resonance spectroscopy; Tn antigen; VVLB4, isolectin B4 from Vicia villosa; Lectin; Tn antigen; O-Glycosylation; mAb, monoclonal antibody; VVLB4, isolectin B4 from Vicia villosa; Monoclonal antibody; Surface plasmon resonance spectroscopy; Cancer
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/S0014-5793(00)01248-5

Using synthetic Tn (GalNAc- O-Ser/Thr) glycopeptide models and a biosensor based on surface plasmon resonance spectroscopy we have determined that isolectin B4 from Vicia villosa (VVLB4) binds to one Tn determinant whereas the anti-Tn monoclonal antibodies 83D4 and MLS128 require at least two Tn residues for recognition. When an unglycosylated amino acid is introduced between the Tn residues, both antibodies do not bind. MLS128 affinity was higher on a glycopeptide with three consecutive Tn residues. These results indicate that Tn residues organized in clusters are essential for the binding of these antibodies and indicate a different Tn recognition pattern for VVLB4.