Novel DNP-KLH protein conjugate surface for sensitive detection of TNT on SPR immunosensor

Praveen Singh, Takeshi Onodera, Yutaka Mizuta, Kiyoshi Matsumoto, Norio Miura, Kiyoshi Toko

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

A nanoscale biosensor chip surface for the detection of trinitrotoluene (TNT) was fabricated using dinitrophenylated-keyhole lympet hemocyanin (DNP-KLH) protein conjugate as ligand supported by an underlying 11-amino 1-undecanethiol hydrochloride (AUT) self-assembled monolayer (SAM) and bis sulfo-succinimidyl suberate (BS3) as crosslinker. Bioactive thin films were fabricated over gold chip via layer-by-layer self-assembly methods. The biomolecular interaction between substrate-specific TNP-glycine-KLH mouse Ig antibody and DNP-KLH conjugate surface was monitored using a surface plasmon resonance-based optical sensor. The quantitation of (TNT) on this bioactive surface was carried out using the solution-based competitive inhibition assay. The DNP-KLH surface biosensor has shown a detection limit of 0.15 ng/ml (150 ppt) and was sensitive up to the 0.005 ng/ml (5 ppt) level for the TNT molecule. This TNT-specific biosensor holds the promise to be one of the most sensitive, fast TNT detecting sensor surfaces under indirect competitive assay format. A 12 s injection pulse of 10 mM glycine-HCl solution was sufficient for the regeneration of the DNP-KLH surface for repeated use. The DNP-KLH sensor platform was checked for its reproducibility and storability.

Original languageEnglish
Pages (from-to)261-273
Number of pages13
JournalSensors and Materials
Volume19
Issue number5
Publication statusPublished - Oct 1 2007

Fingerprint

Trinitrotoluene
Immunosensors
Hemocyanin
trinitrotoluene
proteins
Proteins
bioinstrumentation
Biosensors
glycine
activity (biology)
Glycine
Amino acids
Assays
chips
sensors
Sensors
Optical sensors
hydrochlorides
Surface plasmon resonance
Self assembled monolayers

All Science Journal Classification (ASJC) codes

  • Instrumentation
  • Materials Science(all)

Cite this

Singh, P., Onodera, T., Mizuta, Y., Matsumoto, K., Miura, N., & Toko, K. (2007). Novel DNP-KLH protein conjugate surface for sensitive detection of TNT on SPR immunosensor. Sensors and Materials, 19(5), 261-273.

Novel DNP-KLH protein conjugate surface for sensitive detection of TNT on SPR immunosensor. / Singh, Praveen; Onodera, Takeshi; Mizuta, Yutaka; Matsumoto, Kiyoshi; Miura, Norio; Toko, Kiyoshi.

In: Sensors and Materials, Vol. 19, No. 5, 01.10.2007, p. 261-273.

Research output: Contribution to journalArticle

Singh, P, Onodera, T, Mizuta, Y, Matsumoto, K, Miura, N & Toko, K 2007, 'Novel DNP-KLH protein conjugate surface for sensitive detection of TNT on SPR immunosensor', Sensors and Materials, vol. 19, no. 5, pp. 261-273.
Singh, Praveen ; Onodera, Takeshi ; Mizuta, Yutaka ; Matsumoto, Kiyoshi ; Miura, Norio ; Toko, Kiyoshi. / Novel DNP-KLH protein conjugate surface for sensitive detection of TNT on SPR immunosensor. In: Sensors and Materials. 2007 ; Vol. 19, No. 5. pp. 261-273.
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