TY - JOUR
T1 - Enzyme-linked immunosorbent assay based on light absorption of enzymatically generated aniline oligomer
T2 - Flow injection analysis for 3-phenoxybenzoic acid with anti-3-phenoxybenzoic acid monoclonal antibody
AU - Ishimatsu, Ryoichi
AU - Shimizu, Shinichi
AU - Hongsibsong, Surat
AU - Nakano, Koji
AU - Malasuk, Chacriya
AU - Oki, Yuji
AU - Morita, Kinichi
N1 - Funding Information:
This research in part of monoclonal antibody production was supported by The Thailand Research Fund , Thailand (No. TRG6080007 ) and Chiang Mai University, Chiang Mai , Thailand.
Funding Information:
This research in part of monoclonal antibody production was supported by The Thailand Research Fund, Thailand (No. TRG6080007) and Chiang Mai University, Chiang Mai, Thailand.
Publisher Copyright:
© 2020 Elsevier B.V.
PY - 2020/10/1
Y1 - 2020/10/1
N2 - A flow enzyme-linked immunosorbent assay (ELISA) method based on light absorption by enzymatically generated aniline oligomer in the presence of horseradish peroxidase (HRP), H2O2, and aniline is proposed. Aniline oligomer is rapidly formed through the polymerization reaction via the enzymatic reaction, and its fast reaction rate is beneficial for flow ELISA. An anti-3-phenoxybenzoic acid monoclonal antibody (mAb) was produced by mice, and was used for the flow competitive ELISA for the determination of 3-phenoxybenzoic acid (3PBA), which was performed on an acrylic plate having a Y-shaped channel. ABS resin beads (d = 1 mm) were filled in the channel to increase the surface area for the adsorption of the mAb. A clank-type detection chamber (optical length: 1 cm) made of polydimethylsiloxane (PDMS) containing carbon black, which can significantly decrease light scattering, was fabricated with a 3D printer. The PDMS detection chamber was connected to the outlet of the acrylic flow chip with a tube. A blue LED was used as a light source for the flow ELISA. The inhabitation concentration at 50% and the detection range (absorbance change from 90 to 10%) for the proposed flow competitive ELISA were 0.5 ppm and 0.05–5 ppm, respectively. We also performed the flow competitive ELISA in an artificial and real urine, and no significant matrix effect of the urine samples on the ELISA was found.
AB - A flow enzyme-linked immunosorbent assay (ELISA) method based on light absorption by enzymatically generated aniline oligomer in the presence of horseradish peroxidase (HRP), H2O2, and aniline is proposed. Aniline oligomer is rapidly formed through the polymerization reaction via the enzymatic reaction, and its fast reaction rate is beneficial for flow ELISA. An anti-3-phenoxybenzoic acid monoclonal antibody (mAb) was produced by mice, and was used for the flow competitive ELISA for the determination of 3-phenoxybenzoic acid (3PBA), which was performed on an acrylic plate having a Y-shaped channel. ABS resin beads (d = 1 mm) were filled in the channel to increase the surface area for the adsorption of the mAb. A clank-type detection chamber (optical length: 1 cm) made of polydimethylsiloxane (PDMS) containing carbon black, which can significantly decrease light scattering, was fabricated with a 3D printer. The PDMS detection chamber was connected to the outlet of the acrylic flow chip with a tube. A blue LED was used as a light source for the flow ELISA. The inhabitation concentration at 50% and the detection range (absorbance change from 90 to 10%) for the proposed flow competitive ELISA were 0.5 ppm and 0.05–5 ppm, respectively. We also performed the flow competitive ELISA in an artificial and real urine, and no significant matrix effect of the urine samples on the ELISA was found.
UR - http://www.scopus.com/inward/record.url?scp=85084412829&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85084412829&partnerID=8YFLogxK
U2 - 10.1016/j.talanta.2020.121102
DO - 10.1016/j.talanta.2020.121102
M3 - Article
C2 - 32797869
AN - SCOPUS:85084412829
SN - 0039-9140
VL - 218
JO - Talanta
JF - Talanta
M1 - 121102
ER -