A rapid and highly sensitive immunoassay format for human lipocalin-2 using multiwalled carbon nanotubes.
A sensitive and rapid sandwich immunoassay (IA) was developed for human lipocalin-2 (LCN2) by functionalizing a KOH-treated polystyrene microtiter plate with multiwalled carbon nanotubes (MWCNTs) dispersed in 3-aminoproyltriethoxysilane (APTES). The significantly increased surface area due to the presence of MWCNTs led to a high immobilization density of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) activated anti-LCN2 capture antibodies (Ab). The anti-LCN2 Ab-bound MTPs were stable for 6 weeks when stored in 0.1M PBS, pH 7.4 at 4°C. The IA detects LCN2 from 0.6 to 5120pgmL-1with a limit of detection (LOD) and limit of quantification (LOQ) of 0.9pgmL-1and 6pgmL-1, respectively. The assay offered a ~50-fold lower LOD and ~3-fold faster IA, compared to a commercial sandwich enzyme-linked immunosorbent assay kit.
Institute of Clinical Chemistry and Pathobiochemistry, Klinikum rechts der Isar der Technischen Universität München, Ismaninger Str. 22, Munich d-81675, Germany; Vallo Med Health Care GmbH, Bahnhofstr. 200, Castrop-Rauxel 44579, Germany. Electronic address: sandeep.vashist@vallomed.com.
Sandeep Kumar
SK
Vashist
Innovative Chromatography Group, Irish Separation Science Cluster (ISSC), Department of Chemistry and Analytical, Biological Chemistry Research Facility (ABCRF), University College Cork, Cork, Ireland.
John H T
JHT
Luong
D015374:Biosensing Techniques; D006801:Humans; D007118:Immunoassay; D000071068:Lipocalin-2 / Q000737:chemistry / Q000302:isolation & purification*; D037742:Nanotubes, Carbon / Q000737:chemistry*; D011437:Propylamines / Q000737:chemistry; D012821:Silanes / Q000737:chemistry
C489506:LCN2 protein, human; D000071068:Lipocalin-2; D037742:Nanotubes, Carbon; D011437:Propylamines; D012821:Silanes; C477625:amino-propyl-triethoxysilane
APTES; Human lipocalin-2; Leach-proof immobilization; MWCNTs; Sandwich immunoassay