Development of an NT-ProBNP Assay Reagent Based on High Specific Activity Alkaline Phosphatase CmAP and an Improved Coupling Method

(1) Background: Chemiluminescent enzyme immunoassay (CLEIA) is an efficient analytical method. Alkaline phosphatase (ALP) with high specific activity is the basis for CLEIA to achieve high sensitivity. In this study, a high specific activity <i<Cobetia marina</i< ALP (CmAP) and an improv...
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Gespeichert in:

Autor*in:	Hai-Chao Li [verfasserIn] Xin He [verfasserIn] Shan-Peng Qiao [verfasserIn] Zhen-Ni Liu [verfasserIn] Yu-Zhou Gao [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2020

Schlagwörter:	chemiluminescence enzyme immunoassay (CLEIA) N-terminal prohormone of brain natriuretic peptide (NT-proBNP) antibody coupling

Übergeordnetes Werk:	In: Applied Sciences - MDPI AG, 2012, 10(2020), 23, p 8682
Übergeordnetes Werk:	volume:10 ; year:2020 ; number:23, p 8682

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.3390/app10238682

Katalog-ID:	DOAJ054871026

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520			\|a (1) Background: Chemiluminescent enzyme immunoassay (CLEIA) is an efficient analytical method. Alkaline phosphatase (ALP) with high specific activity is the basis for CLEIA to achieve high sensitivity. In this study, a high specific activity <i<Cobetia marina</i< ALP (CmAP) and an improved coupling method were used to develop an N-terminal pro-B-type natriuretic peptide (NT-proBNP) diagnostic reagent. (2) Methods: The purification method of CmAP was improved and the related enzyme activities were assessed. The enzyme and magnetic beads were coupled only to the Fc region of the detection antibody and the capture antibody, respectively, by using a specially improved method. The NT-proBNP in human serum was assessed. (3) Results: The specific activity of the purified CmAP was found to be 13,133 U/mg. No loss in the enzyme activity was observed after its storage at room temperature for 4 months. The sensitivity of the in vitro diagnostic reagents was found to be 0.58 ng/L. (4) Conclusions: CmAP can be applied as a substitute for the commercial ALP. Analytical parameters indicated that the chemiluminescence diagnostic reagent for NT-proBNP is adequately sensitive and reliable for detecting the serum NT-proBNP, which suggests that both the enzyme and coupling method are suitable for the CLEIA.
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callnumber-first	T - Technology
author	Hai-Chao Li
spellingShingle	Hai-Chao Li misc TA1-2040 misc QH301-705.5 misc QC1-999 misc QD1-999 misc chemiluminescence enzyme immunoassay (CLEIA) misc <i<Cobetia marina</i< alkaline phosphatase (CmAP) misc N-terminal prohormone of brain natriuretic peptide (NT-proBNP) misc antibody coupling misc Technology misc T misc Engineering (General). Civil engineering (General) misc Biology (General) misc Physics misc Chemistry Development of an NT-ProBNP Assay Reagent Based on High Specific Activity Alkaline Phosphatase CmAP and an Improved Coupling Method
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topic_title	TA1-2040 QH301-705.5 QC1-999 QD1-999 Development of an NT-ProBNP Assay Reagent Based on High Specific Activity Alkaline Phosphatase CmAP and an Improved Coupling Method chemiluminescence enzyme immunoassay (CLEIA) <i<Cobetia marina</i< alkaline phosphatase (CmAP) N-terminal prohormone of brain natriuretic peptide (NT-proBNP) antibody coupling
topic	misc TA1-2040 misc QH301-705.5 misc QC1-999 misc QD1-999 misc chemiluminescence enzyme immunoassay (CLEIA) misc <i<Cobetia marina</i< alkaline phosphatase (CmAP) misc N-terminal prohormone of brain natriuretic peptide (NT-proBNP) misc antibody coupling misc Technology misc T misc Engineering (General). Civil engineering (General) misc Biology (General) misc Physics misc Chemistry
topic_unstemmed	misc TA1-2040 misc QH301-705.5 misc QC1-999 misc QD1-999 misc chemiluminescence enzyme immunoassay (CLEIA) misc <i<Cobetia marina</i< alkaline phosphatase (CmAP) misc N-terminal prohormone of brain natriuretic peptide (NT-proBNP) misc antibody coupling misc Technology misc T misc Engineering (General). Civil engineering (General) misc Biology (General) misc Physics misc Chemistry
topic_browse	misc TA1-2040 misc QH301-705.5 misc QC1-999 misc QD1-999 misc chemiluminescence enzyme immunoassay (CLEIA) misc <i<Cobetia marina</i< alkaline phosphatase (CmAP) misc N-terminal prohormone of brain natriuretic peptide (NT-proBNP) misc antibody coupling misc Technology misc T misc Engineering (General). Civil engineering (General) misc Biology (General) misc Physics misc Chemistry
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title	Development of an NT-ProBNP Assay Reagent Based on High Specific Activity Alkaline Phosphatase CmAP and an Improved Coupling Method
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title_full	Development of an NT-ProBNP Assay Reagent Based on High Specific Activity Alkaline Phosphatase CmAP and an Improved Coupling Method
author_sort	Hai-Chao Li
journal	Applied Sciences
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author_browse	Hai-Chao Li Xin He Shan-Peng Qiao Zhen-Ni Liu Yu-Zhou Gao
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author-letter	Hai-Chao Li
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title_sort	development of an nt-probnp assay reagent based on high specific activity alkaline phosphatase cmap and an improved coupling method
callnumber	TA1-2040
title_auth	Development of an NT-ProBNP Assay Reagent Based on High Specific Activity Alkaline Phosphatase CmAP and an Improved Coupling Method
abstract	(1) Background: Chemiluminescent enzyme immunoassay (CLEIA) is an efficient analytical method. Alkaline phosphatase (ALP) with high specific activity is the basis for CLEIA to achieve high sensitivity. In this study, a high specific activity <i<Cobetia marina</i< ALP (CmAP) and an improved coupling method were used to develop an N-terminal pro-B-type natriuretic peptide (NT-proBNP) diagnostic reagent. (2) Methods: The purification method of CmAP was improved and the related enzyme activities were assessed. The enzyme and magnetic beads were coupled only to the Fc region of the detection antibody and the capture antibody, respectively, by using a specially improved method. The NT-proBNP in human serum was assessed. (3) Results: The specific activity of the purified CmAP was found to be 13,133 U/mg. No loss in the enzyme activity was observed after its storage at room temperature for 4 months. The sensitivity of the in vitro diagnostic reagents was found to be 0.58 ng/L. (4) Conclusions: CmAP can be applied as a substitute for the commercial ALP. Analytical parameters indicated that the chemiluminescence diagnostic reagent for NT-proBNP is adequately sensitive and reliable for detecting the serum NT-proBNP, which suggests that both the enzyme and coupling method are suitable for the CLEIA.
abstractGer	(1) Background: Chemiluminescent enzyme immunoassay (CLEIA) is an efficient analytical method. Alkaline phosphatase (ALP) with high specific activity is the basis for CLEIA to achieve high sensitivity. In this study, a high specific activity <i<Cobetia marina</i< ALP (CmAP) and an improved coupling method were used to develop an N-terminal pro-B-type natriuretic peptide (NT-proBNP) diagnostic reagent. (2) Methods: The purification method of CmAP was improved and the related enzyme activities were assessed. The enzyme and magnetic beads were coupled only to the Fc region of the detection antibody and the capture antibody, respectively, by using a specially improved method. The NT-proBNP in human serum was assessed. (3) Results: The specific activity of the purified CmAP was found to be 13,133 U/mg. No loss in the enzyme activity was observed after its storage at room temperature for 4 months. The sensitivity of the in vitro diagnostic reagents was found to be 0.58 ng/L. (4) Conclusions: CmAP can be applied as a substitute for the commercial ALP. Analytical parameters indicated that the chemiluminescence diagnostic reagent for NT-proBNP is adequately sensitive and reliable for detecting the serum NT-proBNP, which suggests that both the enzyme and coupling method are suitable for the CLEIA.
abstract_unstemmed	(1) Background: Chemiluminescent enzyme immunoassay (CLEIA) is an efficient analytical method. Alkaline phosphatase (ALP) with high specific activity is the basis for CLEIA to achieve high sensitivity. In this study, a high specific activity <i<Cobetia marina</i< ALP (CmAP) and an improved coupling method were used to develop an N-terminal pro-B-type natriuretic peptide (NT-proBNP) diagnostic reagent. (2) Methods: The purification method of CmAP was improved and the related enzyme activities were assessed. The enzyme and magnetic beads were coupled only to the Fc region of the detection antibody and the capture antibody, respectively, by using a specially improved method. The NT-proBNP in human serum was assessed. (3) Results: The specific activity of the purified CmAP was found to be 13,133 U/mg. No loss in the enzyme activity was observed after its storage at room temperature for 4 months. The sensitivity of the in vitro diagnostic reagents was found to be 0.58 ng/L. (4) Conclusions: CmAP can be applied as a substitute for the commercial ALP. Analytical parameters indicated that the chemiluminescence diagnostic reagent for NT-proBNP is adequately sensitive and reliable for detecting the serum NT-proBNP, which suggests that both the enzyme and coupling method are suitable for the CLEIA.
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title_short	Development of an NT-ProBNP Assay Reagent Based on High Specific Activity Alkaline Phosphatase CmAP and an Improved Coupling Method
url	https://doi.org/10.3390/app10238682 https://doaj.org/article/54c7779ecf9141bbbfc10e24164116db https://www.mdpi.com/2076-3417/10/23/8682 https://doaj.org/toc/2076-3417
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Analytical parameters indicated that the chemiluminescence diagnostic reagent for NT-proBNP is adequately sensitive and reliable for detecting the serum NT-proBNP, which suggests that both the enzyme and coupling method are suitable for the CLEIA.</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">chemiluminescence enzyme immunoassay (CLEIA)</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a"><i<Cobetia marina</i< alkaline phosphatase (CmAP)</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">N-terminal prohormone of brain natriuretic peptide (NT-proBNP)</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">antibody coupling</subfield></datafield><datafield tag="653" ind1=" " ind2="0"><subfield code="a">Technology</subfield></datafield><datafield tag="653" ind1=" " ind2="0"><subfield code="a">T</subfield></datafield><datafield tag="653" ind1=" " ind2="0"><subfield code="a">Engineering (General). 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Development of an NT-ProBNP Assay Reagent Based on High Specific Activity Alkaline Phosphatase CmAP and an Improved Coupling Method

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