关键词: 细菌性果斑病菌, 单克隆抗体, 制备, 特性鉴定

Abstract:

Four hybridoma cell lines secreting monoclonal antibodies (mAbs) against SD01, namely 6F, 6D, 7E and 4F,
were generated by immunizing BALB/c mice using wild-type strain SD01 of Acidovorax avenae subsp. citrulli (Aac) as
the immunogen. Ascites antibody titers were 1:102 400, 1:102 400, 1:25 600, and 1:51 200, respectively. Ascites were
purified using saturated ammonium sulfate precipitation and Protein-G affinity chromatography. Sodium dodecyl sulfatepolyacrylamide
gelelectrophoresis (SDS-PAGE) analysis showed high antibody purity. The titers of purified mAbs
(2 mg/mL) were 1:12 800, 1:6 400, 1:3 200, and 1:6 400, respectively. The subclass of mAbs was IgG2 a. Indirect enzymelinked
immunosorbent assay (ELISA) showed that four hybridoma cell lines had different binding capacity to eight strains
of Aac and three species of non-Aac plant pathogens. 6F could bind six strains of Aac, 6D and 4F could bind eight strains
of Aac, 7E could bind five strains of Aac. 6F and 4F had cross-reaction with two of the three species of non-Aac plant
pathogens, and 6D and 7E had no cross-reaction with all the three species of non-Aac plant pathogens.

Key words: bacterial fruit blotch, monoclonal antibody, preparation, characterization