TY - JOUR
T1 - Quality control of traditional chinese medicine by monoclonal antibody method
AU - Shang, Ming Ying
AU - Tian, Min
AU - Tanaka, Hiroyuki
AU - Li, Xiao Wei
AU - Cai, Shao Qing
AU - Shoyama, Yukihiro
PY - 2011/3/1
Y1 - 2011/3/1
N2 - In a previous study, we reported the preparation, characterization, variation, specificity, and sensitivity of an anti-aristolochic acid-II (AA-II) monoclonal antibody. The preparation procedure was as follows. AA-II conjugated with bovine serum albumin was used as an antigen for immunizing BALB/c mice. Splenocytes isolated from the immunized mice were fused with an aminopterin-sensitive mouse myeloma cell line to produce hybridoma cells secreting a monoclonal antibody (MAb) against AA-II. The selected MAb was subsequently cloned. Hapten number, isotype, and an estimated dissociation constant (KD) of the secreted MAb were determined. This MAb was used to establish an ELISA method. The linear range was 0.19-13 μg/ml. Anti-AA-II MAb showed extremely high specificity for AA-II, low crossreactivity (CR) against other AAs or aristololactam-I, and negligible CR (<0.5%) toward other natural compounds with different chemical structures. This study describes the successful application of the ELISA method using anti-AA-II MAb to determine AA-II concentration in several crude drugs derived from Aristolochia species. The highest AA-II concentration (2.82 μg/mg) was observed in the stem of A. manshuriensis, followed by that in the fruit of A. contorta (0.81 μg/mg). In case of A. indica, AA-II concentration in the root was higher than that in the aerial parts. These data indicated that the established ELISA method can be used for the quality control of crude drugs derived from Aristolochia plants.
AB - In a previous study, we reported the preparation, characterization, variation, specificity, and sensitivity of an anti-aristolochic acid-II (AA-II) monoclonal antibody. The preparation procedure was as follows. AA-II conjugated with bovine serum albumin was used as an antigen for immunizing BALB/c mice. Splenocytes isolated from the immunized mice were fused with an aminopterin-sensitive mouse myeloma cell line to produce hybridoma cells secreting a monoclonal antibody (MAb) against AA-II. The selected MAb was subsequently cloned. Hapten number, isotype, and an estimated dissociation constant (KD) of the secreted MAb were determined. This MAb was used to establish an ELISA method. The linear range was 0.19-13 μg/ml. Anti-AA-II MAb showed extremely high specificity for AA-II, low crossreactivity (CR) against other AAs or aristololactam-I, and negligible CR (<0.5%) toward other natural compounds with different chemical structures. This study describes the successful application of the ELISA method using anti-AA-II MAb to determine AA-II concentration in several crude drugs derived from Aristolochia species. The highest AA-II concentration (2.82 μg/mg) was observed in the stem of A. manshuriensis, followed by that in the fruit of A. contorta (0.81 μg/mg). In case of A. indica, AA-II concentration in the root was higher than that in the aerial parts. These data indicated that the established ELISA method can be used for the quality control of crude drugs derived from Aristolochia plants.
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U2 - 10.2174/157016311794519983
DO - 10.2174/157016311794519983
M3 - Article
C2 - 21143136
AN - SCOPUS:79951688795
VL - 8
SP - 60
EP - 65
JO - Current Drug Discovery Technologies
JF - Current Drug Discovery Technologies
SN - 1570-1638
IS - 1
ER -