Development of a monoclonal antibody-based enzyme-linked immunosorbent assay for the analysis of glycyrrhizic acid.
Glycyrrhizic acid (GL) is a major active compound of licorice. The specific monoclonal antibody (MAb) (designated as 8F8A8H42H7) against GL was produced with the immunogen GL-BSA conjugate. The dissociation constant (Kd) value of the MAb was approximately 9.96x10(-10) M. The cross reactivity of the MAb with glycyrrhetic acid was approximately 2.6%. The conventional indirect competitive enzyme-linked immunosorbent assay (icELISA) and simplified icELISA adapted with a modified procedure were established using the MAb. The IC50 value and the detect range by the conventional icELISA were 1.1 ng mL-1 and 0.2-5.1 ng mL-1, respectively. The IC50 value and the detect range by the simplified icELISA were 5.3 ng mL-1 and 1.2-23.8 ng mL-1, respectively. The two icELISA formats were used to analyze GL contents in the roots of wild licorice and different parts of cultivated licorice (Glycyrrhiza uralensis Fisch). The results obtained with the two icELISAs agreed well with those of the HPLC analysis. The correlation coefficient was more than 0.98 between HPLC and the two icELISAs. The two icELISAs were shown to be appropriate, simple, and effective for the quality control of raw licorice root materials.
Zhao
Jing
J
College of Agronomy and Biotechnology, China Agricultural University, Beijing, 100094, People's Republic of China.
Li
Gang
G
Wang
Bao-min
BM
Liu
Wei
W
Nan
Tie-gui
TG
Zhai
Zhi-xi
ZX
Li
Zhao-hu
ZH
Li
Qing X
QX
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002851:Chromatography, High Pressure Liquid; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D019695:Glycyrrhizic Acid / Q000032:analysis* / Q000737:chemistry / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D015394:Molecular Structure
D000911:Antibodies, Monoclonal; D019695:Glycyrrhizic Acid