+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Preparation of a specific monoclonal antibody to asiaticoside for the development of an enzyme-linked immunosorbent assay



Preparation of a specific monoclonal antibody to asiaticoside for the development of an enzyme-linked immunosorbent assay



Analyst 136(5): 1013-1017



Asiaticoside (AS), the major active component of Centella asiatica (L.) Urban, is used as a memory enhancer and for wound healing. We have successfully prepared monoclonal antibodies (MAbs) against AS, and developed an enzyme-linked immunosorbent assay (ELISA) system for its determination. AS was conjugated to the carrier protein bovine serum albumin (BSA), which acted as an immunogen. In order to confirm its immunogenicity, the ratio of hapten in the AS-BSA conjugate was determined by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS). After immunization, hybridomas secreting MAbs against AS were produced by fusing splenocytes with the mouse myeloma cell line, SP2/0-Ag14. After the screening, anti-asiaticoside MAb 2B4 was obtained. Weak cross-reactivities occurred with madecassoside (7.08%), but no cross-reactivities were observed with other related triterpenoid glycosides (<0.01%). The assay was suitable for quantitating AS in the range of 0.78 to 50 µg mL(-1). A good correlation of AS concentrations in crude extracts of C. asiatica between ELISA and HPLC methods was obtained (r(2) = 0.999). The contents of AS in various cultivated C. asiatica samples were assayed by the newly established ELISA. The recovery rates of AS in the samples were in the range of 95-103% with coefficients of variation of <10%. The intra- and inter-assay variations were 3.9 and 4.5%, respectively. The ELISA method described should prove useful as an analytical tool for quality control and standardization of medicinal plants and pharmaceutical products containing AS.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 055134493

Download citation: RISBibTeXText

PMID: 21180694

DOI: 10.1039/c0an00868k


Related references

Preparation of monoclonal antibody and development of enzyme-linked immunosorbent assay specific for Escherichia coli O157 in foods. Biomedical and Environmental Sciences 18(4): 254-259, 2005

Development of monoclonal antibody-based ultrasensitive enzyme-linked immunosorbent assay and fluorescence-linked immunosorbent assay for 1-aminohydantoin detection in aquatic animals. Journal of Pharmaceutical and Biomedical Analysis 147: 417-424, 2018

Preparation of monoclonal antibody and development of an indirect competitive enzyme-linked immunosorbent assay for ornidazole detection. Food Chemistry 229: 439-444, 2017

Development of an enzyme-linked immunosorbent assay system for the determination of asymmetric dimethylarginine using a specific monoclonal antibody. Bioscience Biotechnology and Biochemistry 76(2): 400-403, 2012

Development of a specific monoclonal antibody for grouper (Epinephelus guaza) identification by an indirect enzyme-linked immunosorbent assay. Journal of Food Protection 66(5): 886-889, 2003

Development of a Specific Monoclonal Antibody for Grouper (Epinephelus guaza) Identification by an Indirect Enzyme-Linked Immunosorbent Assay. Journal of Food Protection 66(5): 886-889, 2003

Development of specific enzyme-linked immunosorbent antibody assay systems for the detection of chicken immunoglobulins G, M, and A using monoclonal antibodies. Poultry Science 71(2): 302-310, 1992

Development of an indirect competitive enzyme-linked immunosorbent assay (icELISA) using highly specific monoclonal antibody against paclitaxel. Journal of Natural Medicines 67(3): 512-518, 2013

Development of a sandwich enzyme-linked immunosorbent assay for the detection of CD44v3 using exon v3- and v6-specific monoclonal antibody pairs. Journal of Immunological Methods 436: 22-28, 2016

Preparation of an anti-thiamethoxam monoclonal antibody for development of an indirect competitive enzyme-linked immunosorbent assay and a colloidal gold immunoassay. Food and Agricultural Immunology 29(1): 1173-1183, 2018

Preparation of a monoclonal antibody against amantadine and rimantadine and development of an indirect competitive enzyme-linked immunosorbent assay for detecting the same in chicken muscle and liver. Journal of Pharmaceutical and Biomedical Analysis 133: 56-63, 2017

Development of a highly sensitive and specific monoclonal antibody based enzyme-linked immunosorbent assay for the detection of a new β-agonist, phenylethanolamine A, in food samples. Journal of the Science of Food and Agriculture 97(3): 1001-1009, 2017

Development of a highly sensitive and specific monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA) for detection of Sudan I in food samples. Talanta 77(5): 1783-1789, 2009

Development of a bi-specific monoclonal antibody for simultaneous detection of rabbit IgG and horseradish peroxidase: use as a general reagent in immunocytochemistry and enzyme-linked immunosorbent assay. Journal of Histochemistry and Cytochemistry 38(2): 191-198, 1990

Development of a Specific Enzyme-Linked Immunosorbent Assay (ELISA) for the Analysis of the Organophosphorous Pesticide Fenthion in Real Samples Based on Monoclonal Antibody. Analytical Letters 44(9): 1591-1601, 2011