Abstract
Antigen Used for Immunization
Antigen was prepared by sodium nifurstyrenate coupled with carrier protein bovine serum albumin (BSA).
Method of Immunization
Eight- to 10-week-old female Balb/c mice were immunized five times with 0.1 mg of nifurstyrenate-BSA at day 0, 20, 34, 48, and 62.
Parental Cell Line Used for Fusion
Mouse SP2/0 myeloma cells.
Selection and Cloning Procedure
The positive clone was selected and cloned three times using indirect competitive ELISA (icELISA), in which plates were coated with nifurstyrenate-OVA (ovalbumin).
Heavy and Light Chains of Immunoglobulin
ELISA showed that the heavy chain of monoclonal antibody 44A11 is IgG1a subclass; the light chain is kappa (κ).
Specificity
The icELISA method was used to measure the IC50 (the drug concentration corresponding to 50% inhibition) of a series of nitrofuran drugs. The cross-reaction rates of the MAb 44A11 with other nitrofuran drugs were all <0.04%, indicating that it has a good specificity to sodium nifurstyrenate.
Specific Antigen Identified
MAb 44A11 strongly reacted with sodium nifurstyrenate.