Production of Monoclonal Antibodies Against Edwardsiella tarda,and Establishment of Double-antibody Sandwich ELISA Method

doi:10.13560/j.cnki.biotech.bull.1985.2016.07.029

Abstract

Abstract: Using Edwardsiella tarda 2CDM001 as antigen and hybridoma technique,we prepared the monoclonal antibodies（mAbs）8D11,8H4,2F4,18C12,16E2,20D3,19G2,5F7,4E6,7C5 and 18H9 against E. tarda. Among these mAbs,8H4 and 2F4 had the poor specificity,presented varied cross reactions with control strains such as Vibrio damsela（ATCC33539）and Vibrio damsela（ATCC33539）. The other 9 mAbs had favorable specificity and no cross reactions with control strains except E. tarda used in the experiment. The result of detecting E. tarda isolates showed that only 20D3 and 8H4 reacted with all the E. tarda isolates,and the rest of mAbs did not reacted with all the E. tarda isolates. Due to the cross reaction of 8H4 with V. damsela（ATCC33539）,thus 20D3 was selected to establish a double-antibody sandwich ELISA for E. tarda with rabbit polyclonal antibody. The double-antibody sandwich ELISA possessed the strong specificity,and of which the limited detecting concentration reached 5 × 10⁷ CFU /mL. In conclusion,this study expanded the mAbs library for E. tarda,and also provided more reference data and alternative materials for further application of E. tarda mAbs.

Key words: Edwardsiella tarda, monoclonal antibodies, double-antibody sandwich ELISA

YANG Chuan QIN Yi-dan HU Qian LI Qiang. Production of Monoclonal Antibodies Against Edwardsiella tarda,and Establishment of Double-antibody Sandwich ELISA Method[J]. Biotechnology Bulletin, 2016, 32(7): 200-205.

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