Antibody Interaction with a Membrane-Bound Fluorescent Ligand on Synthetic Lipid Vesicles

Robert T. Luedtke, Fred Karush, Robert Luedtke

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

The interaction of membrane-bound ligand with bivalent and monovalent fragments of monoclonal antibody was studied by Fluorescence and precipitation analysis using synthetic lipid vesicles. The ligand Ne-[5-(dimethylamino)-naphthyl-l-sulfonyl]lysine was linked to the hydrophobic anchor dipalmitoylphosphatidylethanolamine and ranged between 0.01 and 1 mol % of the membrane components. The effects of cholesterol on the specific interaction were observed over the range of 0–50 mol %. A precipitation assay was developed to evaluate various factors related to the cross-linking of small unilamellar vesicles by bivalent antibody. The cholesterol content was critical for this process as demonstrated by the increased efficiency of precipitation over the range of 0–40 mol % of this component. Fluorescence analysis yielded the parallel finding of increased accessibility of the ligand to the antibody with greater cholesterol content. Increased surface density of the ligand also was found to enhance the intervesicle interaction. Finally, a comparison of the kinetics by fluorescence analysis of the binding of monovalent and bivalent fragments indicated that the bivalent interaction involved primarily the cross-linking of vesicles in accord with published findings of the interaction of monoclonal antibody with cell membrane antigens.

Original languageEnglish
Pages (from-to)5738-5744
Number of pages7
JournalBiochemistry
Volume21
Issue number23
DOIs
StatePublished - 1 Jan 1982

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Ligands
Membranes
Lipids
Antibodies
Fluorescence
Cholesterol
Monoclonal Antibodies
Unilamellar Liposomes
Cell membranes
Anchors
Lysine
Assays
Cell Membrane
Antigens
Kinetics

Cite this

Luedtke, Robert T. ; Karush, Fred ; Luedtke, Robert. / Antibody Interaction with a Membrane-Bound Fluorescent Ligand on Synthetic Lipid Vesicles. In: Biochemistry. 1982 ; Vol. 21, No. 23. pp. 5738-5744.
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Antibody Interaction with a Membrane-Bound Fluorescent Ligand on Synthetic Lipid Vesicles. / Luedtke, Robert T.; Karush, Fred; Luedtke, Robert.

In: Biochemistry, Vol. 21, No. 23, 01.01.1982, p. 5738-5744.

Research output: Contribution to journalArticle

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