Highly sensitive β-galactosidase detection using streptavidin-display E. coli and lateral flow immunoassay (2023)

Lin, W. Z., Wang, J. P., Ma, I. C., Hsieh, P. C., Hung, Y. J., Hung, C. M. (2023). Highly sensitive β-galactosidase detection using streptavidin-display E. coli and lateral flow immunoassay. Sensors and Actuators, A: Physical, 350, [114114]. https://doi.org/10.1016/j.sna.2022.114114

Lin, Wen Zhi ; Wang, Jun Pei ; Ma, I. Cheng et al. / Highly sensitive β-galactosidase detection using streptavidin-display E. coli and lateral flow immunoassay. In: Sensors and Actuators, A: Physical. 2023 ; Vol. 350.

@article{a485c432054844729b197652f8b8d906,

title = "Highly sensitive β-galactosidase detection using streptavidin-display E. coli and lateral flow immunoassay",

abstract = "Lateral flow immunoassay (LFIA) is a detection method widely used in biomedicine, agriculture, food, and environmental sciences and has the advantages of speed, simplicity, and low cost. However, the poor detection limit of LFIA hinders its application. In this study, we constructed a streptavidin-display E. coli strain to improve the sensitivity of LFIA. Gold nanoparticles (AuNPs) were used as the detected labels and recombinant E. coli binding biotinylated anti-target antibodies served as a signal amplifier. For detection of β-galactosidase, the model protein used in this study, the detection limit was about 5 * 10−15 mol (5 *10−11 M), while that of the conventional LFIA is about 10−12 mol (10−8 M). Having AuNP as the detected label improved LFIA sensitivity 200-fold without sacrificing its advantages and the data interpretation was the same as the traditional LFIA. We further expressed enhanced green fluorescent protein (eGFP) inside the streptavidin-displayed E. coli. Without AuNPs, the fluorescent E. coli acted as a very strong signal, which could be detected by a fluorescence detector, such as a fluorescence microscope. Using the eGFP E. coli as the signal, the detection limit was about 5 * 10−18 mol (5 *10−14 M). This is 2 * 105-fold and 1000-fold better than the AuNP results for the conventional, and proposed method, respectively. However, the method using eGFP is a better fit for lab-use than for point-of-care because of the need for a fluorescence detector and different data interpretation compared with the traditional LFIA. These assays are very promising, especially for rapid screening of proteins as biomarkers.",

keywords = "Gold nanoparticles, Green fluorescent protein, Immunochromatography, Lateral flow immunoassays, Protein detection, Recombinant E. coli",

author = "Lin, {Wen Zhi} and Wang, {Jun Pei} and Ma, {I. Cheng} and Hsieh, {Ping Chun} and Hung, {Yi Jen} and Hung, {Chin Mao} and Hou, {Shao Yi}",

note = "Publisher Copyright: {\textcopyright} 2023 Elsevier B.V.",

year = "2023",

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day = "1",

doi = "10.1016/j.sna.2022.114114",

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volume = "350",

journal = "Sensors and Actuators, A: Physical",

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Lin, WZ, Wang, JP, Ma, IC, Hsieh, PC, Hung, YJ, Hung, CM 2023, 'Highly sensitive β-galactosidase detection using streptavidin-display E. coli and lateral flow immunoassay', Sensors and Actuators, A: Physical, vol. 350, 114114. https://doi.org/10.1016/j.sna.2022.114114

Highly sensitive β-galactosidase detection using streptavidin-display E. coli and lateral flow immunoassay. / Lin, Wen Zhi; Wang, Jun Pei; Ma, I. Cheng et al.
In: Sensors and Actuators, A: Physical, Vol. 350, 114114, 01.02.2023.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - Highly sensitive β-galactosidase detection using streptavidin-display E. coli and lateral flow immunoassay

AU - Lin, Wen Zhi

AU - Wang, Jun Pei

AU - Ma, I. Cheng

AU - Hsieh, Ping Chun

AU - Hung, Yi Jen

AU - Hung, Chin Mao

AU - Hou, Shao Yi

N1 - Publisher Copyright:© 2023 Elsevier B.V.

PY - 2023/2/1

Y1 - 2023/2/1

N2 - Lateral flow immunoassay (LFIA) is a detection method widely used in biomedicine, agriculture, food, and environmental sciences and has the advantages of speed, simplicity, and low cost. However, the poor detection limit of LFIA hinders its application. In this study, we constructed a streptavidin-display E. coli strain to improve the sensitivity of LFIA. Gold nanoparticles (AuNPs) were used as the detected labels and recombinant E. coli binding biotinylated anti-target antibodies served as a signal amplifier. For detection of β-galactosidase, the model protein used in this study, the detection limit was about 5 * 10−15 mol (5 *10−11 M), while that of the conventional LFIA is about 10−12 mol (10−8 M). Having AuNP as the detected label improved LFIA sensitivity 200-fold without sacrificing its advantages and the data interpretation was the same as the traditional LFIA. We further expressed enhanced green fluorescent protein (eGFP) inside the streptavidin-displayed E. coli. Without AuNPs, the fluorescent E. coli acted as a very strong signal, which could be detected by a fluorescence detector, such as a fluorescence microscope. Using the eGFP E. coli as the signal, the detection limit was about 5 * 10−18 mol (5 *10−14 M). This is 2 * 105-fold and 1000-fold better than the AuNP results for the conventional, and proposed method, respectively. However, the method using eGFP is a better fit for lab-use than for point-of-care because of the need for a fluorescence detector and different data interpretation compared with the traditional LFIA. These assays are very promising, especially for rapid screening of proteins as biomarkers.

AB - Lateral flow immunoassay (LFIA) is a detection method widely used in biomedicine, agriculture, food, and environmental sciences and has the advantages of speed, simplicity, and low cost. However, the poor detection limit of LFIA hinders its application. In this study, we constructed a streptavidin-display E. coli strain to improve the sensitivity of LFIA. Gold nanoparticles (AuNPs) were used as the detected labels and recombinant E. coli binding biotinylated anti-target antibodies served as a signal amplifier. For detection of β-galactosidase, the model protein used in this study, the detection limit was about 5 * 10−15 mol (5 *10−11 M), while that of the conventional LFIA is about 10−12 mol (10−8 M). Having AuNP as the detected label improved LFIA sensitivity 200-fold without sacrificing its advantages and the data interpretation was the same as the traditional LFIA. We further expressed enhanced green fluorescent protein (eGFP) inside the streptavidin-displayed E. coli. Without AuNPs, the fluorescent E. coli acted as a very strong signal, which could be detected by a fluorescence detector, such as a fluorescence microscope. Using the eGFP E. coli as the signal, the detection limit was about 5 * 10−18 mol (5 *10−14 M). This is 2 * 105-fold and 1000-fold better than the AuNP results for the conventional, and proposed method, respectively. However, the method using eGFP is a better fit for lab-use than for point-of-care because of the need for a fluorescence detector and different data interpretation compared with the traditional LFIA. These assays are very promising, especially for rapid screening of proteins as biomarkers.

KW - Gold nanoparticles

KW - Green fluorescent protein

KW - Immunochromatography

KW - Lateral flow immunoassays

KW - Protein detection

KW - Recombinant E. coli

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U2 - 10.1016/j.sna.2022.114114

DO - 10.1016/j.sna.2022.114114

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AN - SCOPUS:85145783416

SN - 0924-4247

VL - 350

JO - Sensors and Actuators, A: Physical

JF - Sensors and Actuators, A: Physical

M1 - 114114

ER -

Lin WZ, Wang JP, Ma IC, Hsieh PC, Hung YJ, Hung CM et al. Highly sensitive β-galactosidase detection using streptavidin-display E. coli and lateral flow immunoassay. Sensors and Actuators, A: Physical. 2023 Feb 1;350:114114. doi: 10.1016/j.sna.2022.114114