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| 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 |
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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.
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2020, Vol. 34 
