Binding kinetics of antibody to hapten-doped lipid monolayers as studied by multiple internal reflection fluorescence method
✍ Scribed by Satoshi Tanimoto; Hiromi Kitano
- Book ID
- 103967645
- Publisher
- Elsevier Science
- Year
- 1995
- Tongue
- English
- Weight
- 486 KB
- Volume
- 4
- Category
- Article
- ISSN
- 0927-7765
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✦ Synopsis
Binding processes of an antibody (anti-dinitrophenyl(DNP)-immunoglobulin E) labeled with fluorescein groups (FITC-anti-DNP-IgE) to a hapten-doped lipid monolayer were followed by the multiple internal reflection fluorescence method, because of its usefulness in detecting the adsorption or binding of target molecules at the interface. The hapten-doped lipids used in this work were dinitrophenyl-phosphatidylethanolamine (DNP-DPPE) and dinitrophenylaminocaproyl-phosphatidylethanolamine (DNP-cap-DPPE), and a monolayer containing the hapten-doped lipid was deposited on a poly(methyl methacrylate) cuvette by using the Langmuir-Blodgett technique.
The fluorescence signal increased with the binding of the antibodies, and the initial binding rate was calculated from the slope of the fluorescence signal curve. The initial rate of binding of FITC-anti-DNP-IgE to the hapten on the lipid monolayer surface increased at a low surface density of haptens in the lipid monolayer, and leveled off at a moderate-to-high hapten density. The effect of the spacer length between the hapten group and the lipid layer on the binding rate was also examined. It is confirmed that a long spacer promoted the access of antibodies to the haptens.