NDLI: New Insights into Butyrylcholinesterase Activity Assay: Serum Dilution Factor as a Crucial Parameter

Content Provider	PubMed Central
Author	Joanna, Jońca Żuk, Monika Bartosz, Wasąg Anna, Janaszak-jasiecka Lewandowski, Krzysztof Bartosz, Wielgomas Waleron, Krzysztof Jacek, Jasiecki
Editor	Vrana, Kent E.
Copyright Year	2015
Abstract	Butyrylcholinesterase (BChE) activity assay and inhibitor phenotyping can help to identify patients at risk of prolonged paralysis following the administration of neuromuscular blocking agents. The assay plays an important role in clinical chemistry as a good diagnostic marker for intoxication with pesticides and nerve agents. Furthermore, the assay is also commonly used for in vitro characterization of cholinesterases, their toxins and drugs. There is still lack of standardized procedure for measurement of BChE activity and many laboratories use different substrates at various concentrations. The purpose of this study was to validate the BChE activity assay to determine the best dilution of human serum and the most optimal concentration of substrates and inhibitors. Serum BChE activity was measured using modified Ellman’s method applicable for a microplate reader. We present our experience and new insights into the protocol for high-throughput routine assays of human plasma cholinesterase activities adapted to a microplate reader. During our routine assays used for the determination of BChE activity, we have observed that serum dilution factor influences the results obtained. We show that a 400-fold dilution of serum and 5mM S-butyrylthiocholine iodide can be successfully used for the accurate measurement of BChE activity in human serum. We also discuss usage of various concentrations of dibucaine and fluoride in BChE phenotyping. This study indicates that some factors of such a multicomponent clinical material like serum can influence kinetic parameters of the BChE. The observed inhibitory effect is dependent on serum dilution factor used in the assay.
Related Links	http://dx.doi.org/10.1371/journal.pone.0139480
Starting Page	139480
File Format	PDF
ISSN	19326203
e-ISSN	19326203
Journal	PLoS ONE
Issue Number	10
Volume Number	10
Language	English
Publisher	Public Library of Science
Publisher Date	2015-10-01
Access Restriction	Open
Rights Holder	Public Library of Science
Subject Keyword	Biochemistry, Genetics and Molecular Biology(all) Agricultural and Biological Sciences(all) Medicine(all) Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

New Insights into Placozoan Sexual Reproduction and Development

New Insights into [FeFe] Hydrogenase Activation and Maturase Function

New Insights into Samango Monkey Speciation in South Africa

Structural Insights into Cellulolytic and Chitinolytic Enzymes Revealing Crucial Residues of Insect β-N-acetyl-D-hexosaminidase

New Insights into the Evolution of Metazoan Tyrosinase Gene Family

Chromosomal Passports Provide New Insights into Diffusion of Emmer Wheat

Correction: New Insights into Samango Monkey Speciation in South Africa

New Insights into the Binding and Catalytic Mechanisms of Bacillus thuringiensis Lactonase: Insights into B. thuringiensis AiiA Mechanism

New Insights into the Phylogeny and Molecular Classification of Nicotinamide Mononucleotide Deamidases

New Insights into Butyrylcholinesterase Activity Assay: Serum Dilution Factor as a Crucial Parameter

Similar Documents

New Insights into Placozoan Sexual Reproduction and Development

New Insights into [FeFe] Hydrogenase Activation and Maturase Function

New Insights into Samango Monkey Speciation in South Africa

Structural Insights into Cellulolytic and Chitinolytic Enzymes Revealing Crucial Residues of Insect β-N-acetyl-D-hexosaminidase

New Insights into the Evolution of Metazoan Tyrosinase Gene Family

Chromosomal Passports Provide New Insights into Diffusion of Emmer Wheat

Correction: New Insights into Samango Monkey Speciation in South Africa

New Insights into the Binding and Catalytic Mechanisms of Bacillus thuringiensis Lactonase: Insights into B. thuringiensis AiiA Mechanism

New Insights into the Phylogeny and Molecular Classification of Nicotinamide Mononucleotide Deamidases

New Insights into Butyrylcholinesterase Activity Assay: Serum Dilution Factor as a Crucial Parameter