Preparation and Immunological Characterization of Monoclonal Antibody against Furacillin Metabolite
XU Dongmei, LI Yaying, GENG Haibo, LI Yujing
1.Department of Food and Drug Engineering, Shi Jia Zhuang University of Applied Technology, Shijiazhuang 050081, China; 2.Department of Physical Education, Hebei University of Science and Technology, Shijiazhuang 050018, China; 3.Discipline Inspection, Supervision and Audit Division, Hebei Jiaotong Vocational and Technical College, Shijiazhuang 050035, China
Abstract：In order to establish an immunological method for the detection of the furacillin metabolite semicarbazide (SEM), a conjugate of SEM with bovine serum albumin (BSA) was synthesized by the carbodiimide method and was used as an immunogen to immunize BALB/c mice for the preparation of a monoclonal antibody against SEM (SEM mAb) by polyethylene glycol (PEG)-mediated cell fusion. SEM mAb was purified by caprylic acid-ammonium sulfate precipitation. Finally, the immunological characteristics of the monoclonal antibody including titer, sensitivity, specificity and affinity were identified. The results showed that the monoclonal antibody SEM-3D9 was successfully prepared. The serum titer of immunized mice was 1:5 × 106, the sensitivity IC50 was 0.42 μg/L, and the antibody belonged to IgG1 subtype with an affinity constant Ka of 2.1 × 108 L/mol. Its cross-reaction rate with 2-NP-3-amino-2-oxazolidinone and 2-nitrophenyl-5-morpholine-methyl-3-amino-2-oxazolidinone analogues was less than 0.01%, and its cross-reaction rate with 2-nitrophenyl-1-aminohydantoin was 0.016 8%, indicating good specificity.
2020, Vol. 34 
