title: 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.
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin, Heilongjiang, PR China.
authors.forename: Ming
authors.initials: M
authors.lastname: Xu
authors.forename: Dongbo
authors.initials: D
authors.lastname: Sun
authors.forename: Jinlong
authors.initials: J
authors.lastname: Li
authors.forename: Shiwen
authors.initials: S
authors.lastname: Xu
mesh_terms: D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000276:immunology*; D000915:Antibody Affinity; D000918:Antibody Specificity; D001717:Birds / Q000235:genetics / Q000276:immunology*; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D006825:Hybridomas / Q000276:immunology; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology*; D051151:Selenoprotein W / Q000235:genetics / Q000276:immunology*
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D011994:Recombinant Proteins; D051151:Selenoprotein W; D007544:Isopropyl Thiogalactoside