The purpose of this study was to generate monoclonal antibodies (mAb) specific for chicken interleukin-4 (ChIL-4) and develop a capture ELISA for the detection of ChIL-4. Five mAbs against ChIL-4 were generated by immunizing Balb/c mice with recombinant ChIL-4 (rChIL-4). As determined in ELISA, immunofluorescent assay and western blotting, each mAb reacted specifically with rChIL-4 derived from baculovirus and Escherichia coli (E. coli) expression systems. In order to identify a suitable pair of mAbs for the development of ChIL-4 capture ELISA, mAbs were tested as both capture and revealing antibodies. To achieve the highest sensitivity of the capture ELISA, mAb 20C9 was used as the capture antibody and biotinylated mAb 16D8 was used as the revealing antibody, which allowed the detection of ChIL-4 low to 500 pg/ml. The specificity of ELISA was verified using rChIL-4 derived from baculovirus expression system and irrelevant proteins as control. The results showed that this ELISA is suitable to detect rChIL-4 in different forms and would provide a sensitive tool to measure the in vitro release of ChIL-4 in birds.

To Cite This Article: Dai H, Chen JH,  Shen Y, Zhou X, Xu ZZ, Chen X and Jiao XA, 2018. Production of monoclonal antibodies against chicken interleukin-4 and development of a capture enzyme-linked immunosorbent assay. Pak Vet J, 38(3): 286-290. http://dx.doi.org/10.29261/pakvetj/2018.066