Immobilization of proteins onto novel, reactive polypyrrole-coated polystyrene latex particles (original) (raw)
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Polypyrrole core/polyacrolein shell latex for protein immobilization
Colloid & Polymer Science, 1995
The redox polymerization of pyrrole, with ferric chloride as oxidant, carried out in the presence of polyvinylpyrrolidone (PVP), yielded polypyrrole latex particles. The polypyrrole latex was used, as seed, for the radical polymerization of acrolein. The resulting pol~,pyrrole core/polyacrolein shell tat~x (poly(P/A)) was suitable for immobilization of up to 11 mg of human serum albumin (HSA) and/or 33 mg of human gamma globulin (?G) per t g of latex particles.
Colloid and Polymer Science, 2004
Over the last two decades inherently conducting polymers (ICPs) have attracted a great deal of interest owing to their remarkable physical and chemical properties, such as inherent conductivity, redox, and acid-base properties [1]. Polypyrrole (PPy) is one of the most studied ICPs because it offers reasonably high conductivity and has fairly good environmental stability with regard to air and water. In addition, it is easily synthesized in high yield via oxidative polymerization at room temperature in various common solvents, including water [2]. However, PPy is invariably obtained as a black precipitate which is insoluble in all common solvents.
Langmuir, 2004
Novel ester-functionalized polypyrrole-silica nanocomposite particles were prepared by oxidative copolymerization of pyrrole and N-succinimidyl ester pyrrole (50/50% initial concentrations), using FeCl3 in the presence of ultrafine silica nanoparticles (20 nm diameter). The N-succinimidyl ester pyrrole monomer was prepared in aqueous solution using 1-(2-carboxyethylpyrrole) and N-hydroxysuccinimide in the presence of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide. The resulting nanocomposites (N-succinimidyl ester polypyrrole-silica) are raspberry-shaped agglomerates of silica sol particles "glued" together by the insoluble poly(pyrrole-co-N-succinimidyl pyrrole). The N-succinimidyl ester polypyrrole-silica particles were characterized in terms of their size, density, copolymer content, and polydispersity. Scanning electron microscopy and disk centrifuge sedimentometry confirmed that the nanocomposite particles had narrow size distributions. X-ray photoelectron spectroscopy analysis indicated a silica-rich surface and a high surface concentration of N-succinimidyl ester groups. These nanoparticles exhibited good long-term dispersion stability. The chemical stability of the ester functions in aqueous media after several weeks of storage was monitored by FTIR spectroscopy. The functionalized nanocomposites were tested as bioadsorbents of human serum albumin (HSA). The very high amount of immobilized HSA determined by UV-visible spectroscopy is believed to be due to covalent binding. Incubation of the HSA-grafted nanocomposite with anti-HSA resulted in immediate flocculation, an indication that they are alternative candidates for visual diagnostic assays. * Corresponding
Polymers for Advanced Technologies, 2003
It has been well-established that nanomaterials provide a robust framework into which two or more functional moieties can be integrated to offer multifunctional and synergetic applications. We report here the facile synthesis and systematical investigation of the luminomagnetic core-shell nanoparticles (NPs) with the magnetic Fe 3 O 4 core coated with a silica shell incorporating fluorescent [Ru(bpy) 3 ] 2+ . The luminomagnetic NPs were monodisperse and spherical in shape with a diameter of 60 ± 10 nm. The luminomagnetic NPs possessed not only the desirable optical signature of Ru(bpy) 3 2+ but also the distinctive magnetic profile of Fe 3 O 4 , where a strong red-orange emission and the super-paramagnetic characteristics with the saturation magnetization values ca. 10 emu/g were observed for the luminomagnetic NPs. As revealed by Alamar blue assay and flow cytometry analysis, the Fe 3 O 4 NPs decrease the cell viability of HepG2 by ca. 10%, while an increase by ca. 10% on HepG2 cell proliferation was revealed after the silica shell was coated onto Fe 3 O 4 NPs, suggesting that the silica shell serves as a protective layer to increase the biocompatibility of the luminomagnetic NPs. Confocal laser scanning microscopy, transition electron microscopy and magnetic resonance (MR) images confirmed that the luminomagnetic NPs can enter into the interiors of HepG2 cells without damage, highlighting their capabilities for simultaneous optical fluorescence imaging and T2 MR imaging. Taking advantage of versatility of silica shell towards different surface modification protocols, the luminomagnetic NPs were successfully functionalized with epidermal growth factor receptor (EGFR) antibody for HepG2 cell recognition. All the results illustrated that the luminomagnetic NPs should be a potential candidate for future cancer diagnosis and therapy.
Interfacial physicochemical properties of functionalized conducting polypyrrole particles
Polymer, 2005
Polypyrrole-coated polystyrene latex particles bearing reactive N-succinimidyl ester functional groups (PS-PPyNSE 75 ) were prepared by the in situ copolymerization of pyrrole 1 and the active N-succinimidyl ester-functionalized pyrrole 2 (pyrroleNSE), with initial 1:2 fractions of 25:75 (%) in the presence of sterically stabilized polystyrene (PS) latex particles. PS particles were prepared by dispersion polymerization leading to particles having a diameter of 600G10 nm. The PS-PPyNSE 75 particles were characterized in terms of surface morphology and chemical composition. Surface analysis of the colloidal materials by X-ray photoelectron spectroscopy (XPS) and scanning electron microscopy (SEM) indicated a substantial coating of PS by the reactive conducting copolymer. Infrared spectroscopy permitted to detect pyrroleNSE repeat units at the surface of the particles indicating that 1 and 2 did indeed copolymerise.
Efficient Immobilization of Proteins by Modification of Plate Surface with Polystyrene Derivatives
Analytical Biochemistry, 1997
stance, in the case of ELISA, 3 reactivities of antibodies Immobilization of proteins on microplate wells by to antigens can be measured by immobilizing antigens simple adsorption (e.g., for ELISA) is convenient, but onto microplates. Similarly, carbohydrate binding acit can be inefficient, especially if proteins are hydrotivities and specificities of lectins can be measured philic or small in size. This problem was alleviated by quantitatively using immobilized lectins and labeled the use of polyvinylbenzyl lactonoylamide (PVLA). ligands. However, this conventional method is much PVLA is strongly adsorbed to the hydrophobic well too dependent on the hydrophobicity or the size of prosurface, and its lactonamide part can be oxidized with tein because immobilization onto polystyrene-like surperiodate to generate aldehydo groups. Proteins are faces relies mostly on hydrophobic interaction. When then immobilized covalently to the aldehydo groups a protein is hydrophilic or small, immobilization is usuby reductive amination under mild conditions. Using ally inefficient which results in large amounts of prothis method, henceforth termed the PVLA method, altein being required for immobilization. Therefore, it is kaline phosphatase (AP) was immobilized to midesirable to have a more universal method of immobilicroplates six-to sevenfold greater than by simple adzation that is less dependent on the nature of proteins. sorption (as measured by activity). Similarly, the Polyvinylbenzyl lactonoylamide (PVLA) is a polystyactivity of immobilized mannose-binding protein A rene derivative with pendant lactonoylamide chains (MBP-A) was 4-to 8-fold higher by the PVLA method (1). This polymer was originally synthesized as an asiathan by simple adsorption. The PVLA-coated plates loglycoprotein mimic, and it was found that the PVLAneeded as little as 200 ng of MBP-A per well to have a coated culture dishes promoted the differentiated funcsufficient amount of MBP-A immobilized for the meation and proliferation of hepatocyte culture (2, 3). We surement of binding of 125 I-labeled mannosylated bodescribe in this paper a new method of protein immobivine serum albumin (125 I-Man-BSA), but unmodified plates required as much as 20 mg/well MBP-A to obtain lization that utilizes the hydrophobic nature of polystythe same response. Recommended conditions for the rene-like structure and the hydrophilic nature of the PVLA method are 40 ml of 2 mg/ml of PVLA for coating, carbohydrate part of PVLA. The polystyrene-like struc-1 mM NaIO 4 for the generation of the aldehydo groups, ture of PVLA should result in a strong and stable coatand a 2-h reductive amination at 37ЊC between pH 8 ing of the microplate surface. The hydrophilic carbohyand 9 for the protein ligation.
Biotechnology and Bioengineering, 1989
Reactive polymers have been prepared by copolymerizing N-isopropyl acrylamide (NIPAM) with N-acryloxysuccinimide (NASI) or glycidyl methacrylate (GMA). The amino groups of ligands could react with the residues of NASI or GMA and the polymers could be precipitated by temperature and/or salinity variation, since they contained the NIPAM residues. As a model, p-aminobenzamidine, a trypsin inhibitor, was attached to the polymers to form water-soluble macroligands, capable of selectively binding trypsin from a trypsin-chymotrypsin solution. After precipitation of the macroligand-trypsin complex, followed by dissociation, approximately 82% trypsin was isolated. The NIPAM-GMA copolymer was also reacted with immunogammaglobulin (IgG) and alkaline phosphatase (AP). It was demonstrated that the IgG bearing polymer was able to bind protein A and the whole complex was precipitable. The reactive polymer was also used for direct innmobilization of AP which was active in repeated reactions.
Protein immobilization on formylated polystyrene supports
Reactive and Functional Polymers, 2000
Immobilization of nine various proteins at cross-linked polyformylstyrene shell on 0.63-cm molded polystyrene beads intended for solid phase immunoassay has been investigated. The pH value of the protein solution and NaCl concentration 2 substantially influence the density of the immobilized protein (DIP). The maximum values of DIP (up to 104 mg / cm ) were obtained in pH 5 pI 0.1 M phosphate buffer and in 1 M NaCl, pH 3-4. The DIP decreased with the increase in the proteins molecular weight at a constant initial protein concentration.