Synthesis of artificial chlorogenic acid antigen and preparation of polyclonal antibodies / 中草药

ABSTRACT

Objective: To synthesize artificial chlorogenic acid (CHA) antigen, to prepare CHA polyclonal antibody, and to lay the foundation for the preparation of immune chip for allergic component analysis. Methods: CHA-bovine serum albumin (CHA-BSA) and CHA-ovalbumin (CHA-OVA) were synthesized by carbodiimide 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 determination, the CHA was successfully conjugated with BSA. The serum antibody titer was 1128 000 with the linear range of 15.6-250 ng/mL, R2 = 0.995, and Y = -0.10 lnX+1.158. No cross-reaction was detected with glycyrrhizic acid, parietic acid and so on, which were commonly used in the combination with CHA. Conclusion: The preparation of multi-clonal antibody with good sensitivity and specificity against CHA is successful, which provides a foundation for the trace detection and allergy research of CHA.