Abstract
Antigen Used for Immunization
Succinic acid mono-[bis-(4-chloro-phenyl)-methyl] ester.
Method of Immunization
Female BALB/c mice (8 weeks old) were immunized subcutaneously with 30 μg of DCBH-BSA conjugate emulsified in complete Freund's adjuvant. Mice were given subsequent two injections with the immunogen emulsified in incomplete Freund's adjuvant at 2-week intervals. Mouse selected as spleen donor for hybridoma production was administered intravascularly with 50 μg of immunogen in 100 μL of sterile PBS (pH 7.2).
Parental Cell Line Used for Fusion
P3-X63Ag8.653 myeloma cells.
Selection and Cloning Procedure
Three days after intravenous boosting, mouse spleen was aseptically collected for cell fusion using HAT as a selection medium. Supernatant was screened for specific antibodies by non-competitive indirect ELISA. The corresponding positive wells were selected and clones were selected by limiting dilution assay. The most sensitive clone was expanded in 10% FBS/IMDM.
Heavy and Light Chains of Immunoglobulin
IgG1 kappa chain.
Specificity
Specificity of produced MAb was evaluated by competitive indirect ELISA.
Specific Antigen Identified
Dicofol.