Comparison of Candidate Pairs of Hydrolytic Enzymes for Spectrophotometric-dual-enzyme-simultaneous-assay

Spectrophotometric-dual-enzyme-simultaneous-assay (SDESA) for enzyme-linked-immunosorbent-assay (ELISA) of two components in one well is a patented platform when a special pair of labels is accessible. With microplate readers, alkaline phosphatase on 4-nitro-1-naphthylphosphate (4NNPP) served as lab...

Full description

Saved in:
Bibliographic Details
Published in:Analytical Sciences 2015/05/10, Vol.31(5), pp.421-427
Main Authors: LIU, Hongbo, YUAN, Mei, YANG, Xiaolan, HU, Xiaolei, LIAO, Juan, DANG, Jizheng, XIE, Yanling, PU, Jun, LI, Yuanli, ZHAN, Chang-Guo, LIAO, Fei
Format: Article
Language:English
Subjects:
4-nitro-1-naphthylphosphate
Absorbance
acetylcholinesterase
Acetylcholinesterase - metabolism
Alkaline phosphatase
Alkaline Phosphatase - metabolism
Analytical Chemistry
Arylsulfatases - metabolism
Chemistry
ELISA
Enzyme-Linked Immunosorbent Assay - methods
Enzyme-linked-immunosorbent-assay
Equal channel angular pressing
Hydrolysis
Labels
Naphthalenes - chemistry
Naphthalenes - metabolism
Nitro Compounds - chemistry
Nitro Compounds - metabolism
Nitrophenols - chemistry
Nitrophenols - metabolism
Pseudomonas aeruginosa
Pseudomonas aeruginosa - enzymology
Pseudomonas aeruginosa arylsulfatase
Readers
spectrophotometric-dual-enzyme-simultaneous-assay
Spectrophotometry
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Spectrophotometric-dual-enzyme-simultaneous-assay (SDESA) for enzyme-linked-immunosorbent-assay (ELISA) of two components in one well is a patented platform when a special pair of labels is accessible. With microplate readers, alkaline phosphatase on 4-nitro-1-naphthylphosphate (4NNPP) served as label A; Pseudomonas aeruginosa arylsulfatase (PAAS) and acetylcholinesterase (AChE) on their substrates derived from 4-nitrophenol/analogue served as candidate label B, and were compared for SDESA with an engineered alkaline phosphatase of Eschrichia coli (ECAP). For SDESA, the interference from overlapped absorbance was corrected based on linear additivity of absorbance to derive initial rates reflected by absorbance change at 450 nm for ECAP and at 405 nm for PAAS or AChE, after the correction of spontaneous hydrolysis. For SDESA with ECAP, AChE already had sufficient activity in an optimized buffer; PAAS was more favorable for substrate stability and product absorbance except for lower activity. Therefore, PAAS engineered for sufficient activity plus alkaline phosphatase is absorbing for ELISA via SDESA.
ISSN:0910-6340
1348-2246
DOI:10.2116/analsci.31.421