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Ligand modification of Pluronic F108 for use in immobilized metal affinity separation of bio-macromolecules

Thesis (MSc)--Stellenbosch University, 2002. / ENGLISH ABSTRACT: Inthis work we aim to put into place a system to separate or immobilise biomacromolecules
by means ofimmobilised transition metal ions like nickel(II) or
copper(II). Although the concept of immobilised metal affmity chromatography
(IMAC) has been around since the early 1960's, the metal ions were always
immobilised by covalent modification of the support matrix. Recently the concept of
IMAC was applied to membranes, and again the metal ion was immobilised by
covalent modification of the membrane surface. Inthis study we covalently modified
the support matrix by attaching a linear, EDTA type ligand to the hydroxy end groups
of a tri-block copolymer (polyethylene oxide (PEO)m = 129 - polypropylene oxide
(PPO)n = 56 - polyethylene oxide (PEO)m= 129), Pluronic® F108. The middle block of
this polymer, which is hydrophobic, will non-covalently adsorb onto the membrane
surface through hydrophobic interaction. The hydrophilic outer blocks, with the
ligand modified end groups, will associate with the aqueous substrate exposing the
chelated metal ion for interaction with the bio-macromolecules. This affords a system
which is recyclable, without replacing the membranes, simply by stripping the expired
ligand modified-polymer and adsorbing fresh polymer.
A series of model ligands and their complexes were synthesised and characterised, to
study the coordination of the ligand around the metal ions. The model compounds
were also essential in characterising the final product - the ligand modified Pluronic.
Finally the ligand modified Pluronic was tested for its metal binding capabilities.
This was done in aqueous solution by qualitatively comparing the UV-VIS spectra of
the modified Pluronic with that of the model ligands and complexes. The spectra
indicate that metal coordination does take place. / AFRIKAANSE OPSOMMING: In die studie het ons beoog om 'n sisteem daar te stel vir die skei en immobilisering
van bio-makromolekules deur middel van oorgangsmetale soos nikkel(II) en
koper(II). Alhoewel die beginsel van geïmmobiliseerde Metaal-ioon Affmiteits
Chromatografie (IMAC) reeds sedert die vroeë 1960's bekend is, is die metaal ione
geïmmobiliseer deur kovalente modifikasie van die draermaterial. Onlangs is die
beginsel van IMAC uitgebrei na membrane en ook hierin is die metaalione
geïmmobiliseer deur kovalente modifikasie van die membraanoppervlaktes. In die
projek het ons die draermateriaal kovalent gemodifiseer deur 'n lineêre EDT A-tipe
ligand te koppel aan die hidroksie eindgroepe van 'n tri-blok ko-polimeer (poli-etileen
oksied (PEO)m = 129 - poli-propileen oksied (PPO)n = 56 - poli-etileen oksied (PEO)m =
129), Pluronic® FI08. Die middelste blok van die polimeer, wat hidrofobies is, sal nie-kovalent
aan die membraan oppervlakte adsorbeer d.m. v hidrofobiese interaksie. Die
hidrofiliese buite blokke, met die ligand-gemodifiseerde eindgroepe, sal assosieer met
die waterige substraat en die metal ioon blootstel vir interaksie met die biomakromolekules.
Dit stel dus 'n sisteem in plek wat herbenut kan word, sonder om
die membrane te vervang, deur eenvoudig die ligand-gemodifiseerde polimeer wat
verval het te stroop en te vervang met nuwe polimeer.
'n Reeks modelligande en hul komplekse was gesintetiseer en gekarakteriseer om die
koördinasie van die ligande rondom die metaal ione te bestudeer. Dié model
verbindings was van groot belang in die karakterisering van die finale produk - die
ligand-gemodifiseerde Pluronic.
Laastens is die ligand-gemodifiseerde Pluronic getoets vir sy metaal bindings
vermoeë. Dit is gedoen deur die UV-VIS spectra van die gemodifiseerde Pluronic
kwalitatiefte vergelyk met die spectra van die modelligande en komplekse, in
waterige oplossings. Die spectra dui aan dat metaalbinding wel plaasvind.

Identiferoai:union.ndltd.org:netd.ac.za/oai:union.ndltd.org:sun/oai:scholar.sun.ac.za:10019.1/52918
Date03 1900
CreatorsVan Kralingen, Leon
ContributorsAdendorff, H. J. A., Bredenkamp, M. W., Stellenbosch University. Faculty of Science. Dept. of Chemistry & Polymer Science.
PublisherStellenbosch : Stellenbosch University
Source SetsSouth African National ETD Portal
Languageen_ZA
Detected LanguageEnglish
TypeThesis
Format92 p. : ill.
RightsStellenbosch University

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