ABSTRACT
Immunogenic antigen (spinosin-BSA) and coating antigen (spinosin-OVA) of spinosin were synthesized by sodium periodate oxidation method. UV scanning analysis method showed that these two spinosins were successfully conjugated with carrier protein and the coupling ratio was 17 and 13.7, respectively. Meanwhile, when immunized by spinosin-BSA，the mice can produce anti-spinosin antibodies with the high titer (1:32 000),specificity (IC₅₀ 211.6 μg·L⁻¹) and low cross-reaction rate measured by ELISA tests. The artificial antigen of spinosin was successfully synthesized, which can be applied for preparation of monoclonal antibodies and establishment of appropriate immune method.
ABSTRACT
Immunogenic antigen (jujuboside A-BSA) and coating antigen (jujuboside A-OVA) of jujuboside A were synthesized by sodium periodate oxidation method for the first time. Jujuboside A artificial antigen was confirmed by matrix-assisted laser desorption ionization/time-of-flight mass spectrometry (MALDI-TOF-MS). The titer and specificity of the antibody in serum of immunized mice were detected by enzyme-linked immunosorbent assay (ELISA). The corrected relation curve of inhibition rate showed that the antibody against Jujuboside A obtained from immunized mice could bind to jujuboside A and the titer was up to 1∶4 000. The jujuboside A artificial antigen was synthesized, which can be used further to preparation of monoclonal antibody and the pharmacokinetics study of jujuboside A in laboratory animals.
ABSTRACT
Objective: To synthesize and identify the artificial antigen of loganin for the first time, and provide a foundation for the preparation of specific monoclonal antibody and establishment of immunoassay method. Methods: Sodium periodate oxidation method was used to synthesize immunogenic antigen (loganin-BSA) and coating antigen (loganin-OVA) of loganin. Whether loganin was conjugated with BSA and OVA or not was confirmed by matrix-assisted laser desorption ionization/time-of-flight mass spectrometry (MALDI-TOF-MS). The titer and specificity of the antibody in serum of immunised mice were detected by enzyme-linked immunosorbent assay (ELISA). Results: The results of MALDI-TOF-MS indicated that loganin was conjugated to BSA. The antibody against loganin obtained from immunised-mice could bind to loganin and the titer was up to 1:40 000. Conclusion: The artificial antigen of loganin was synthesized, which can be used further in the preparation of monoclonal antibody, application in quality control of Chinese materia medica and the pharmacokinetic study of loganin in laboratory animals.
ABSTRACT
OBJECTIVE: To synthesize conjugates as artificial complete antigen and coated antigen of icariin to establish an enzyme linked immunosorbent assay (ELISA).
ABSTRACT
Objective: To synthesize the artificial antigen of ginsenoside Rg1-bovine serum albumin (Rg1-BSA) and the artificial coated antigen of ginsenoside Rg1-polylysine (Rg1-PLL), and to provide the basis for the preparation of monoclonal antibody (MAb) and the establishment of immunoassay method. Methods: Rg1-BSA and Rg1-PLL were synthesized by sodium periodate oxidation method. The characterization of the synthesis was examined by UV spectrometry and TLC method. The titer and specificity of the antibody in serum of immunised mice were detected by indirect enzyme-linked immunosorbent assay (I-ELISA) and indirect competitive enzyme-linked immunosorbent assay (I-CELISA), respectively. Results: According to the UV and TLC, the Rg1 was successfully conjugated with BSA and PLL. I-ELISA and IC-ELISA methods were developed using Rg1-PLL. The anti-Rg1 antibody obtained from immunized mice could bind to Rg1 specially and the titer was up to 1:80000. Conclusion: The artificial immunogen Rg1-BSA and coated antigen Rg1-PLL are successfully synthesized, which could be used to prepare the MAb of Rg1 and establish the immunoassay method.