dc.contributor.author Henry, SM
dc.contributor.author Rodionov, I
dc.date.accessioned 2011-10-12T02:32:22Z
dc.date.available 2011-10-12T02:32:22Z
dc.date.copyright 2011
dc.date.issued 2011-10-12
dc.identifier.citation KODE Biotech Technical Bulletin, number 20110928, pp. 1-10
dc.identifier.uri http://hdl.handle.net/10292/2241
dc.description.abstract The FSL-RFG(Maleimide) FSL Construction Kit is for use in creating Function-Spacer-Lipid (FSL) constructs for use in non-covalent cell-surface modification/engineering of cellular membranes, viral particles, liposomes, or other surfaces [1-10]. FSL-RFG(Mal) is one of several FSL constructs with Reactive Functional Groups (RFG); with this construct having maleimide as its Function group. The semi-rigid Spacer in this molecule is constructed via modified hexapeptide unit (Gly-Gly-Ida)2 coupling to both amino groups of ethylenediamine and has been designed to ensure accessibility for target binding/external interactions and proper presentation of functional peptides at a cell or virion surface as well as imparting good solubility to the construct. Electrostatic repulsion forces of spacer’s anionic groups probably favor uniform distribution of the incorporated constructs on the membrane surface [11]. The diacyl phospholipid derived from unsaturated fatty acids is a prerequisite for spontaneous incorporation into cell membranes. This FSL-RFG(Maleimide) FSL Construction Kit cat # 960819-1-R&D (includes a detailed procedure and contains reagents sufficient for one FSL preparation on a milligram scale from cysteine-containing peptides (Figure 1), proteins or any other thiols of biological interest. The effective synthetic approach is based on the well-known Michael nucleophilic addition to maleimides, which react fast and selectively with SH-groups in the pH range 6.5-7.5 producing stable thioether linkages completely stable at physiological conditions [12-15]. The reaction half-life between millimolar concentrations of maleimide and thiol is estimated to be of the order of few seconds [14,15]; but more complex and heavy molecules of biochemical interest interact somewhat slower even when applied in 10-fold excess and durations of at least 2 hours are recommended [16]. The protocol described here is optimized for this kit using FSL-RFG(Mal) with generic peptides and addresses problems which may be encountered if purification of completed FSL constructs is required.
dc.publisher AUT University
dc.relation.ispartofseries number #20110928, pp.1 - 10
dc.rights Auckland University of Technology (AUT) encourages public access to AUT information and supports the legal use of copyright material in accordance with the Copyright Act 1994 (the Act) and the Privacy Act 1993. Unless otherwise stated, copyright material contained on this site may be in the intellectual property of AUT, a member of staff or third parties. Any commercial exploitation of this material is expressly prohibited without the written permission of the owner.
dc.title FSL-RFG(Maleimide) FSL construction kit
dc.type Commissioned Report
dc.rights.accessrights OpenAccess

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