Gum Arabic coated magnetic nanoparticles with affinity ligands specific for antibodies.
A novel magnetic support based on gum Arabic (GA) coated iron oxide magnetic nanoparticles (MNP) has been endowed with affinity properties towards immunoglobulin G (IgG) molecules. The success of the in situ triazine ligand synthesis was confirmed by fluorescence assays. Two synthetic ligands previously developed for binding to IgG, named as ligand 22/8 (artificial Protein A) and ligand 8/7 (artificial Protein L) were immobilized on to MNPs coated with GA (MNP_GA). The dimension of the particles core was not affected by the surface functionalization with GA and triazine ligands. The hydrodynamic diameters of the magnetic supports indicate that the coupling of GA leads to the formation of larger agglomerates of particles with about 1 microm, but the introduction of the triazine ligands leads to a decrease on MNPs size. The non-functionalized MNP_GA bound 28 mg IgG/g, two times less than bare MNP (60 mg IgG/g). MNP_GA modified with ligand 22/8 bound 133 mg IgG/g support, twice higher than the value obtained for ligand 8/7 magnetic adsorbents (65 mg/g). Supports modified with ligand 22/8 were selected to study the adsorption and the elution of IgG. The adsorption of human IgG on this support followed a Langmuir behavior with a Q(máx) of 344 mg IgG/g support and K(a) of 1.5 x 10(5) M. The studies on different elution conditions indicated that although the 0.05 M citrate buffer (pH 3) presented good recovery yields (elution 64% of bound protein), there was occurrence of iron leaching at this acidic pH. Therefore, a potential alternative would be to elute bound protein with a 0.05 M glycine-NaOH (pH 11) buffer.
Batalha
Iris L
IL
REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal.
Hussain
Abid
A
Roque
A C A
AC
D000327:Adsorption; D000818:Animals; D000906:Antibodies / Q000737:chemistry*; D000915:Antibody Affinity; D005290:Ferric Compounds / Q000737:chemistry; D006170:Gum Arabic / Q000737:chemistry*; D006801:Humans; D008024:Ligands; D008280:Magnetics; D053768:Metal Nanoparticles / Q000737:chemistry*; D015394:Molecular Structure; D010316:Particle Size; D012685:Sepharose / Q000737:chemistry; D017550:Spectroscopy, Fourier Transform Infrared; D014227:Triazines / Q000737:chemistry
D000906:Antibodies; D005290:Ferric Compounds; D008024:Ligands; D014227:Triazines; C000499:ferric oxide; D006170:Gum Arabic; D012685:Sepharose