Preparation and Enzyme-Linked Immunosorbent Assay of Monoclonal Antibody Against Ampicillin

Abstract

Ampicillin (AMP) was coupled with mcKLH by the method of EDC intermediate and injected into Balb/c mice as immunogen. The hybridomas were obtained by fusing mouse myeloma cells SP2/0 with splenocytes from the mice immunized with AMP-KLH. The chromosomal numbers were 97-104, the results indicated that the subclass of the McAb was IgG2a, the titer of ascitic fluid was 2×10 6 , the affinity constant was 3×10 -10 mol/L, the molecular weight was 154KD. The cross-reactivities of McAb to other antibiotics were below 0.01%. The linear detecting range of calibration curves to detect AMP was 0.5 ng/mL-100 ng/mL. The formular was Y = 0.0569X+0.1308, R 2 = 0.9948. The limit of detection was 0.3 ng/mL.