Monoclonal Antibodies Against Avian Selenoprotein W

Abstract

The Sel-W gene encoding avian selenoprotein W was cloned into the vector pGEX-6p-1 with GST tag. The recombinant protein Sel-W was expressed in Escherichia coli BL21 (DE3) after induction with 1 mM IPTG. Female 10-week-old BALB/c mice were immunized with the purified Sel-W recombinant protein emulsified in Freund's adjuvant. Two monoclonal antibodies against the Sel-W, 1B8 and 4H5, were produced by lymphocyte hybridoma technique. Subtypes of the MAbs 1B8 and 4H5 were all IgM, and their ascitic fluid titers were 1:2800 and 6400, respectively. In specificity, the MAbs 1B8 and 4H5 showed positive reaction with the recombinant Sel-W protein with GST tag and the synthesized Sel-W protein, and could not react with the GST tag and the recombinant Sel-N protein. In sensitivity, the detection limits of the MAbs 1B8 and 4H5 for the recombinant Sel-W protein and the synthesized Sel-W protein were 39 ng/mL and 52 ng/mL, respectively. These data suggest that the MAb 1B8 and 4H5 will have a potential use for detection and function analysis of the avian Sel-W.

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