Attachment of Water-Soluble Proteins to the Surface of (Magnetizable) Phospholipid Colloids via NeutrAvidin-Derivatized Phospholipids
✍ Scribed by Marcel De Cuyper; Michael Hodenius; Zulmira G.M. Lacava; Ricardo B. Azevedo; Maria de Fátima da Silva; Paulo Cesar Morais; Maria Helena Andrade Santana
- Publisher
- Elsevier Science
- Year
- 2002
- Tongue
- English
- Weight
- 118 KB
- Volume
- 245
- Category
- Article
- ISSN
- 0021-9797
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✦ Synopsis
The present work describes the incorporation of a functionalized phospholipid derivative into the phospholipid bilayer of both classical small unilamellar vesicles and recently developed magnetoliposomes, resulting in unique biocolloid structures onto which peripheral water-soluble enzymes can be immobilized on the surfaces. In the first part of this work, a synthesis protocol is outlined for a universal membrane anchor for water-soluble proteins. Dioleoylphosphatidylethanolamine-N-dodecanyl was used as the starting lipid molecule. After activation of the terminal -COOH group, alpha,omega-diamino-poly(ethylene glycol), used as a hydrophilic, flexible spacer arm, was coupled covalently. Subsequently, NeutrAvidin was bound, after blocking the free -NH(2) groups with citraconic anhydride. In the second part, the resulting lipid-NeutrAvidin derivative was incorporated into small unilamellar vesicles comprised of dimyristoylphosphatidylglycerol. FPLC with Superdex 200 as the column matrix clearly showed that biotinylated alkaline phosphatase, which served as a representative model of water-soluble proteins, was attached to the vesicles. Furthermore, magnetoliposomes, constructed of the same type of phospholipid molecules, were presented as interesting colloids to assess the degree of enzyme immobilization in a rapid and elegant manner. Potential applications that can emerge from this study are briefly discussed.