A reliable aptamer array prepared by repeating inkjet-spotting toward on-site measurement.
A preparation protocol is proposed for a reliable aptamer array utilizing an ink-jet spotter. We accumulated streptavidin and biotinylated-aptamer in this order on a biotinylated-polyethylene glycol-coated gold substrate to prepare an aptamer array. The aptamer array was prepared with an alternate spotting structure where each aptamer spot was placed between reference spots formed with blocking solution thus suppressing contamination from neighboring spots during the blocking and washing processes. Four aptamer spots were prepared in a small area of 1×4.8mm(2) with five reference spots made of blocking solution. We evaluated the thrombin binding ability of the spotted aptamer array using a multi-analysis surface plasmon resonance sensor. We prepared a disposable capillary-driven flow chip designed for on-site measurement (Miura et al., 2010) with our aptamer array and detected thrombin from phosphate-buffered saline at concentrations of 50ngmL(-1) and 1μgmL(-1) (equivalent to 1.35 and 27nM, respectively). A correlation was observed between the refractive index shift and thrombin concentration. This implies that our array preparation protocol meets the requirement for the preparation of a one-time-use chip for on-site measurement.
NTT Device Technology Laboratories, Nippon Telegraph and Telephone Corporation, 3-1 Morinosato Wakamiya, Atsugi-shi, Kanagawa 243-0198, Japan.
Suzuyo
S
Inoue
NTT Device Technology Laboratories, Nippon Telegraph and Telephone Corporation, 3-1 Morinosato Wakamiya, Atsugi-shi, Kanagawa 243-0198, Japan. Electronic address: seyama.michiko@lab.ntt.co.jp.
Michiko
M
Seyama
NTT Device Technology Laboratories, Nippon Telegraph and Telephone Corporation, 3-1 Morinosato Wakamiya, Atsugi-shi, Kanagawa 243-0198, Japan.
Toru
T
Miura
NTT Device Technology Laboratories, Nippon Telegraph and Telephone Corporation, 3-1 Morinosato Wakamiya, Atsugi-shi, Kanagawa 243-0198, Japan.
Tsutomu
T
Horiuchi
NTT Device Technology Laboratories, Nippon Telegraph and Telephone Corporation, 3-1 Morinosato Wakamiya, Atsugi-shi, Kanagawa 243-0198, Japan.
Yuzuru
Y
Iwasaki
NTT Device Technology Laboratories, Nippon Telegraph and Telephone Corporation, 3-1 Morinosato Wakamiya, Atsugi-shi, Kanagawa 243-0198, Japan.
Jun-Ichi
JI
Takahashi
NTT Device Technology Laboratories, Nippon Telegraph and Telephone Corporation, 3-1 Morinosato Wakamiya, Atsugi-shi, Kanagawa 243-0198, Japan.
Katsuyoshi
K
Hayashi
NTT Device Technology Laboratories, Nippon Telegraph and Telephone Corporation, 3-1 Morinosato Wakamiya, Atsugi-shi, Kanagawa 243-0198, Japan.
Emi
E
Tamechika
D052157:Aptamers, Nucleotide / Q000737:chemistry*; D019921:Biotinylation; D004867:Equipment Design; D006046:Gold / Q000737:chemistry; D006801:Humans; D056656:Lab-On-A-Chip Devices; D011092:Polyethylene Glycols / Q000737:chemistry; D020349:Surface Plasmon Resonance / Q000295:instrumentation* / Q000379:methods; D013917:Thrombin / Q000032:analysis*
D052157:Aptamers, Nucleotide; C081676:thrombin aptamer; D011092:Polyethylene Glycols; D006046:Gold; D013917:Thrombin
Aptamer array; Blood analysis; Capillary driven flow; Inkjet spotter; Surface plasmon resonance (SPR)