Determination of conjugation efficiency of antibodies and proteins to the superparamagnetic iron oxide nanoparticles by capillary electrophoresis with laser-induced fluorescence detection
2003 (English)In: Journal of nanoparticle research, ISSN 1388-0764, E-ISSN 1572-896X, Vol. 5, no 02-jan, 137-146 p.Article in journal (Refereed) Published
The method based on capillary electrophoresis with laser-induced fluorescence detection (CE/LIF) was developed for determination of magnetic iron oxide nanoparticles (hydrodynamic diameters of 100 nm) functionalized with molecules containing primary amino groups. The magnetic nanoparticles with carboxylic or aminopropyltrimethoxysilane groups at their surface were conjugated to the model proteins ( bovine serum albumin, BSA; streptavidin or goat anti-rabbit immunoglobulin G, IgG) using carbodiimide as a zero-length cross-linker. The nanoparticle-protein conjugates ( hydrodynamic diameter 163 - 194 nm) were derivatized with naphthalene-2,3- dicarboxaldehyde reagent and separated by CE/LIF with a helium - cadmium laser ( excitation at 442 nm, emission at 488 nm). The separations were carried out by using a fused-silica capillary ( effective length 48 cm, inner diameter 75 mum) and 100 mM sodium borate buffer ( pH 9.2), the potential was 30 kV. The detection limit for BSA-conjugate was 1.3 pg/10 nl, i.e. about 20 amol. The present method provides an efficient and fast tool for sensitive determination of the efficacy of biomolecular functionalization of magnetic nanoparticles. The CE/LIF technique requires only negligible sample volumes for analysis, which is especially suitable for controlling the process of preparation of functionalized nanoparticles with unique properties aimed to be used for diagnostic or therapeutic purposes.
Place, publisher, year, edition, pages
2003. Vol. 5, no 02-jan, 137-146 p.
capillary electrophoresis, laser-induced fluorescence, magnetic nanoparticles, albumin, streptavidin, IgG, micellar electrokinetic chromatography, zone-electrophoresis, biomedical applications, gene-expression, amino-acids, separation, particles, systems, mr
IdentifiersURN: urn:nbn:se:kth:diva-22615ISI: 000183704000017OAI: oai:DiVA.org:kth-22615DiVA: diva2:341313
QC 201005252010-08-102010-08-10Bibliographically approved