Covalent immunochemical membrane labeling of viable cells with K698-T708, a simian virus 40 tumor antigen-derived peptide.

Abstract

The process of covalent immunochemical linking of viable cell membranes with a Simian Virus 40 (SV40) tumor antigen-derived undecapeptide, K(698)PPTPPPEPET(708) (KT), is described. The principle applied was the reaction of the lysine residue, K 698, of the undecapeptide with the succinimidyl moiety of a heterobifunctional linker molecule, N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP) or sulfosuccinimidyl(4-iodo-acetyl)aminobenzoate (sulfo-SIAB). Thereby, upon release of N-hydroxy-succinimide, the rest of the linker molecule reacts covalently with the epsilon-NH2 group of lysine. Upon release of pyridyl-2-thion or hydrogen iodide, respectively, the second reactive moiety of the linker is then ready to form a covalent bond with SH-groups of cell membrane compounds. As a result, KT is covalently linked onto the cell membrane by an -SS- or an -S-bond, respectively. Binding is prevented by treatment of the candidate cells with iodoacetamide, an SH-reactive compound. This artificial cell membrane epitope can be demonstrated by surface immunofluorescence and by binding to immunomagnetic beads loaded with PAb1605, a KT-specific monoclonal antibody. Quantitation by cytofluorimetry shows some 10(4) KT molecules bound per cell, a number that is in the range of the number of SV40 tumor antigen molecules of genuine SV40-transformed mammalian cells.

• Authors: R Hess, P Rau, M Schwab, S Paetzold, M Kuther, M Obert, H Agostini, C Haessler, D G Braun, G Brandner
• Journal: Peptide research (Pept Res) Vol. 7 Issue 3 Pg. 146-52 ( 1994) ISSN: 1040-5704 [Print] United States
• PMID: 7521699 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antigens, Polyomavirus Transforming
• Cross-Linking Reagents
• Epitopes
• K698-T708 undecapeptide
• Oligopeptides
• Vaccines

Topics

• Animals
• Antigens, Polyomavirus Transforming (chemistry)
• Cell Membrane (chemistry)
• Cross-Linking Reagents
• Epitopes (chemistry)
• Flow Cytometry
• Fluorescent Antibody Technique
• Lymphoid Tissue (cytology)
• Mice
• Neoplasms, Experimental (therapy)
• Oligopeptides (chemistry, immunology)
• Vaccines (chemistry)