Skip to Content
Merck
  • Development of surface plasmon resonance imaging biosensors for detection of ubiquitin carboxyl-terminal hydrolase L1.

Development of surface plasmon resonance imaging biosensors for detection of ubiquitin carboxyl-terminal hydrolase L1.

Analytical biochemistry (2014-10-15)
Anna Sankiewicz, Piotr Laudanski, Lech Romanowicz, Adam Hermanowicz, Wiesława Roszkowska-Jakimiec, Wojciech Debek, Ewa Gorodkiewicz
ABSTRACT

We have developed a new method for highly selective determination of the ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1) concentration using a surface plasmon resonance imaging (SPRI) technique and two different biosensors. UCH-L1 was captured from a solution by immobilized specific rabbit monoclonal antibody or specific LDN-57444 inhibitor due to formation of receptor-UCH-L1 complex on the biosensor surface. The analytically useful dynamic response range of both biosensors is between 0.1 and 2.5ng/ml. The detection limit is 0.06ng/ml for the biosensor with antibody and 0.08ng/ml for the biosensor with inhibitor. Biosensors based on both antibody and inhibitor were found to be suitable for quantitative determination of the UCH-L1 and exhibit good tolerance to the potential interferents. Both biosensors gave comparable results in the range of 0 to 0.20ng/ml for plasma samples and 0.30 to 0.49ng/ml for cerebrospinal fluid samples. To validate the new methods, comparative determination of UCH-L1 by the commercial enzyme-linked immunosorbent assay (ELISA) kit was performed. In general, in terms of UCH-L1 concentration, a good correlation between SPRI and ELISA was found. The developed biosensors can be used successfully for the determination of UCH-L1 in body fluids.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Acetic acid-12C2, 99.9 atom % 12C
Sigma-Aldrich
Sodium chloride solution, 0.85%
Sigma-Aldrich
Magnesium chloride solution, 0.1 M
Sigma-Aldrich
Hydrogen chloride solution, 3 M in cyclopentyl methyl ether (CPME)
Sigma-Aldrich
Sodium acetate, SAJ first grade, ≥98.0%
Supelco
Ethanol solution, certified reference material, 2000 μg/mL in methanol
Sigma-Aldrich
Sodium chloride, random crystals, 99.9% trace metals basis
Sigma-Aldrich
Sodium hydroxide, SAJ first grade, ≥95.0%
Sigma-Aldrich
Sodium carbonate, anhydrous, powder, 99.999% trace metals basis
Sigma-Aldrich
Acetic acid, natural, ≥99.5%, FG
Sigma-Aldrich
Potassium chloride, 99.999% trace metals basis
Sigma-Aldrich
Sodium chloride, 99.999% trace metals basis
Sigma-Aldrich
Acetic acid, ≥99.5%, FCC, FG
Sigma-Aldrich
Sodium Acetate Anhydrous, >99%, FG
Sigma-Aldrich
Magnesium chloride solution, BioUltra, Molecular Biology, ~1 M in H2O
Sigma-Aldrich
Sodium carbonate, BioUltra, anhydrous, ≥99.5% (calc. on dry substance, T)
Sigma-Aldrich
Potassium chloride, AnhydroBeads, −10 mesh, 99.99% trace metals basis
Supelco
ISA (ionic strength adjustment solution: 1 M KCl), 1 M KCl
Sigma-Aldrich
Magnesium chloride solution, BioUltra, Molecular Biology, ~0.025 M in H2O
Sigma-Aldrich
Acetic acid, suitable for luminescence, BioUltra, ≥99.5% (GC)
Sigma-Aldrich
Sodium acetate, anhydrous, BioUltra, suitable for luminescence, Molecular Biology, ≥99.0% (NT)
Supelco
Potassium chloride solution, ~3 M KCl, saturated with silver chloride
Supelco
Sodium chloride, reference material for titrimetry, certified by BAM, >99.5%
Supelco
Hydrogen chloride – methanol solution, ~1.25 m HCl (T), derivatization grade (GC derivatization), LiChropur
Sigma-Aldrich
Sodium chloride solution, BioUltra, Molecular Biology, ~5 M in H2O
Supelco
Hydrogen chloride – ethanol, ~1.25 M HCl, derivatization grade (GC derivatization), LiChropur
Sigma-Aldrich
Potassium chloride, BioUltra, Molecular Biology, ≥99.5% (AT)
Supelco
Acetic acid, analytical standard
Supelco
Potassium chloride solution, BioUltra, ~3 M in H2O
Sigma-Aldrich
Sodium acetate solution, BioUltra, Molecular Biology, ~3 M in H2O