Fab fragments imprinted SPR biosensor for real-time human immunoglobulin G detection
(2011) In Biosensors and Bioelectronics 28(1). p.97-104- Abstract
F(ab) fragments imprinted surface plasmon resonance (SPR) chip was prepared for the real-time detection of human immunoglobulin G (IgG). In order to attach polymerization precursor on SPR chip, the SPR chip surface was modified with allyl mercaptan. F(ab) fragments of the IgG molecules were prepared by papain digestion procedure and collected by fast protein liquid chromatography (FPLC) system using Hi-Trap_r Protein A FF column. The collected F(ab) fragments were complexed with histidine containing specific monomer, N-methacryloyl-l-histidine methyl ester (MAH). Molecular imprinted polymeric nanofilm was prepared on SPR chip in the presence of ethylene glycol dimethacrylate and 2-hydroxyethylmethacrylate. The template molecules, F(ab)... (More)
F(ab) fragments imprinted surface plasmon resonance (SPR) chip was prepared for the real-time detection of human immunoglobulin G (IgG). In order to attach polymerization precursor on SPR chip, the SPR chip surface was modified with allyl mercaptan. F(ab) fragments of the IgG molecules were prepared by papain digestion procedure and collected by fast protein liquid chromatography (FPLC) system using Hi-Trap_r Protein A FF column. The collected F(ab) fragments were complexed with histidine containing specific monomer, N-methacryloyl-l-histidine methyl ester (MAH). Molecular imprinted polymeric nanofilm was prepared on SPR chip in the presence of ethylene glycol dimethacrylate and 2-hydroxyethylmethacrylate. The template molecules, F(ab) fragments, were removed from the polymeric nanofilm using 1M NaCl solution (pH: 7.4, phosphate buffer system). The molecular imprinted SPR chip was characterized by contact angle, atomic force microscopy and Fourier transform infrared spectroscopy. By the real-time IgG detection studies carried out using aqueous IgG solutions in different concentrations, the kinetics and isotherm parameters of the molecular imprinted SPR chip-IgG system were calculated. To show selectivity and specificity of the molecular imprinted SPR chip, competitive kinetic analyses were performed using bovine serum albumin (BSA), IgG, F(ab) and F(c) fragments in singular and competitive manner. As last step, IgG detection studies from human plasma were performed and the measured IgG concentrations were well matched with the results determined by enzyme-linked immunosorbent assay (ELISA). The results obtained with the molecular imprinted SPR chip were well fitted to Langmuir isotherm and the detection limit was found as 56 ng/mL. In the light of the results, we can conclude that the proposed molecular imprinted SPR chip can detect IgG molecules from both aqueous solutions and complex natural samples.
(Less)
- author
- Ertürk, Gizem LU ; Uzun, Lokman ; Tümer, M Aşkın ; Say, Rıdvan and Denizli, Adil
- publishing date
- 2011
- type
- Contribution to journal
- publication status
- published
- keywords
- Biosensing Techniques, Humans, Immunoglobulin Fab Fragments, Immunoglobulin G, Mathematics, Molecular Imprinting, Surface Plasmon Resonance, Journal Article
- in
- Biosensors and Bioelectronics
- volume
- 28
- issue
- 1
- pages
- 97 - 104
- publisher
- Elsevier
- external identifiers
-
- pmid:21802938
- scopus:80052342524
- ISSN
- 1873-4235
- DOI
- 10.1016/j.bios.2011.07.004
- language
- English
- LU publication?
- no
- id
- d1492367-a48c-4c1e-947e-1b410d5042eb
- date added to LUP
- 2018-01-25 13:57:09
- date last changed
- 2024-05-27 06:15:33
@article{d1492367-a48c-4c1e-947e-1b410d5042eb,
abstract = {{<p>F(ab) fragments imprinted surface plasmon resonance (SPR) chip was prepared for the real-time detection of human immunoglobulin G (IgG). In order to attach polymerization precursor on SPR chip, the SPR chip surface was modified with allyl mercaptan. F(ab) fragments of the IgG molecules were prepared by papain digestion procedure and collected by fast protein liquid chromatography (FPLC) system using Hi-Trap_r Protein A FF column. The collected F(ab) fragments were complexed with histidine containing specific monomer, N-methacryloyl-l-histidine methyl ester (MAH). Molecular imprinted polymeric nanofilm was prepared on SPR chip in the presence of ethylene glycol dimethacrylate and 2-hydroxyethylmethacrylate. The template molecules, F(ab) fragments, were removed from the polymeric nanofilm using 1M NaCl solution (pH: 7.4, phosphate buffer system). The molecular imprinted SPR chip was characterized by contact angle, atomic force microscopy and Fourier transform infrared spectroscopy. By the real-time IgG detection studies carried out using aqueous IgG solutions in different concentrations, the kinetics and isotherm parameters of the molecular imprinted SPR chip-IgG system were calculated. To show selectivity and specificity of the molecular imprinted SPR chip, competitive kinetic analyses were performed using bovine serum albumin (BSA), IgG, F(ab) and F(c) fragments in singular and competitive manner. As last step, IgG detection studies from human plasma were performed and the measured IgG concentrations were well matched with the results determined by enzyme-linked immunosorbent assay (ELISA). The results obtained with the molecular imprinted SPR chip were well fitted to Langmuir isotherm and the detection limit was found as 56 ng/mL. In the light of the results, we can conclude that the proposed molecular imprinted SPR chip can detect IgG molecules from both aqueous solutions and complex natural samples.</p>}},
author = {{Ertürk, Gizem and Uzun, Lokman and Tümer, M Aşkın and Say, Rıdvan and Denizli, Adil}},
issn = {{1873-4235}},
keywords = {{Biosensing Techniques; Humans; Immunoglobulin Fab Fragments; Immunoglobulin G; Mathematics; Molecular Imprinting; Surface Plasmon Resonance; Journal Article}},
language = {{eng}},
number = {{1}},
pages = {{97--104}},
publisher = {{Elsevier}},
series = {{Biosensors and Bioelectronics}},
title = {{Fab fragments imprinted SPR biosensor for real-time human immunoglobulin G detection}},
url = {{http://dx.doi.org/10.1016/j.bios.2011.07.004}},
doi = {{10.1016/j.bios.2011.07.004}},
volume = {{28}},
year = {{2011}},
}