Evaluation of 2-Methacryloyloxyethyl Phosphorylcholine Polymeric Nanoparticle for Immunoassay of C-Reactive Protein Detection
- 1 April 2004
- journal article
- research article
- Published by American Chemical Society (ACS) in Analytical Chemistry
- Vol. 76 (9) , 2649-2655
- https://doi.org/10.1021/ac035321i
Abstract
To prepare novel 2-methacryloyloxyethyl phosphorylcholine (MPC)−polymeric nanoparticle (MPC−PNP), water-soluble amphiphilic phospholipid polymer, poly [MPC-co-n-butyl methacrylate (BMA)-co-p-nitrophenyloxycarbonyl poly(ethylene glycol) methacrylate (MEONP) (PMBN)], which has active ester groups for bioconjugation on the side chains, was synthesized. MPC−PNP was prepared by a solvent evaporation technique where the poly(l-lactic acid) was used as core and PMBN was applied as an emulsifier and a surface modifier under systematical design of well-arranged phospholipids polar groups in its surface. Characteristics for MPC−PNP were thoroughly investigated with dynamic light scattering, electrophoresis light scattering, X-ray photoelectron spectroscopy, and field emission scanning electron microscopy measurements. Through a protein adsorption test, the phosphorylcholine group on the surface of MPC−PNPs, which had their active ester groups substituted by glycine, were shown to suppress the nonspecific adsorption of bovine serum albumin. These particles were used for C-reactive protein (CRP) detection, where anti-CRP monoclonal antibodies were immobilized on the MPC−PNP using the active ester group, while the remaining active ester groups were thoroughly reacted with glycine. The detection limit about serum-free CRP in the calibration curve was shown to extend from 0.01 to 10 mg/dL when anti-CRP antibody immobilized MPC−PNP was used for serum-free CRP detection. This compares favorably with measurement using polystyrene nanoparticles that were shown to detect from 0.1 to 10 mg/dL by an immunoagglutination technique. Also, for the detection of CRP in serum, MPC−PNP was shown to give the same calibration curve explained by the efficient suppression of nonspecific binding. Furthermore, denaturation of immobilizing anti-CRP antibody on the MPC−PNP hardly occurred despite increasing the temperature. It is concluded that MPC−PNP is unique due to the design of its interfacial properties, also it will perform well in a diagnostic immunoassay because of its optimized material properties.Keywords
This publication has 23 references indexed in Scilit:
- Structure of Water in the Vicinity of Phospholipid Analogue Copolymers As Studied by Vibrational SpectroscopyLangmuir, 2003
- Biomembrane surfaces as models for polymer design: the potential for haemocompatibilityPublished by Elsevier ,2003
- Production of Protein-Loaded Polymeric Microcapsules by Compressed CO2 in a Mixed SolventIndustrial & Engineering Chemistry Research, 2001
- In Situ Assembly of Colloidal Particles into Miniaturized BiosensorsLangmuir, 1999
- Synthesis, structure and surface dynamics of phosphorylcholine functional biomimicking polymersBiomaterials, 1998
- Why do phospholipid polymers reduce protein adsorption?Journal of Biomedical Materials Research, 1998
- Protein adsorption from human plasma is reduced on phospholipid polymersJournal of Biomedical Materials Research, 1991
- The serological diagnosis of human hydatid disease by time-resolved fluoroimmunoassayJournal of Infection, 1991
- Latex piezoelectric immunoassay: Detection of agglutination of autibody-bearing latex using a piezoelectric quartz crystal.CHEMICAL & PHARMACEUTICAL BULLETIN, 1990
- The Fluid Mosaic Model of the Structure of Cell MembranesScience, 1972