Aim: To synthesize two kinds of amoxicillin artificial antigen and to establish hybridoma cell lines secreting monoclonal antibody against amoxillin.
Methods: Amoxicillin were respectively coupled with ovalbumin(OVA) and bovine serum albumin(BSA) by using N-hydroxysuc-cinimide active ester (NHS) method to prepared the immunogen and detection antigen. Molar ratio of conjugates were detected by ultraviolet spectrum analysis. BALB/c mice was immunized with AMX-OVA. The hybridomas were obtained by fusing mouse myeloma cells Sp2/0 with splenocytes from the mice immunized with AMX-OVA. The aseites with mAb were purified by octanoic acid-saturated ammonium sulphate method. Isotype was determined by Mouse Isotype kit (Bio-RadLabs.).
Results: UV spectra shows overlay features between a small molecular compound and carrier protein. Five hybridoma cell strains secreting specific mAb against amoxicillin were obtained and the five Amb belong to IgG1 kappa isotype. The limit of detection of AMX was 0.25 ng/mL and but there is mild degree of cross-reactions with ampicillin with same structure of β-lactam ring.
Conclusion: The anti-AMX Amb could be used to develop immunoassay for detection of residues.