title: [Secretable eukaryotic expression of recombinant chicken Iglambda and preparation of monoclonal antibodies against chicken Iglambda].
abstract: OBJECTIVE
  To construct the eukaryotic expression vector for chicken Iglambda light chain, to express it on COS7 cells and to prepare the monoclonal antibodies against chicken Iglambda.
  
  
  METHODS
  The cDNA of chicken Iglambda light chain with signal peptide sequence was amplified and then inserted into eukaryotic expression plasmid pcDNA3 after double enzyme cutting. The constructed recombinant vector was transfected into COS7 cells by lipofectamin and the secretable eukaryotic expression of chicken Iglambda light chain was verified by Western blot. The monoclonal antibodies (mAbs) against chicken Iglambda light chain were prepared by immunizing BALB/c mice with 2 x 10(6) chicken B cells and by cell fusion technology.
  
  
  RESULTS
  The eukaryotic expression vector was successfully constructed. Western blot demonstrated that chicken Iglambda light chain existed in the cultural supernatant. The hybridoma lines secreting anti-Iglambda mAbs were screened by indirect ELISA. The specific reactivity between anti-Iglambda mAbs and recombinant chicken Iglambda light chain was detected by Western blot.
  
  
  CONCLUSIONS
  The secreted recombinant chicken Iglambda light chain and anti-Iglambda mAbs provide a basis for further study of the functions of chicken Iglambda.
authors.affiliation: Academy of Agricultural Sciences in Henan, Zhengzhou 450002, China. angela9922@sina.com
authors.forename: Hong
authors.initials: H
authors.lastname: Zhang
authors.forename: Gai-ping
authors.initials: GP
authors.lastname: Zhang
authors.forename: Jin-gang
authors.initials: JG
authors.lastname: Zhang
authors.forename: Yan-yan
authors.initials: YY
authors.lastname: Yang
authors.forename: Ming-lin
authors.initials: ML
authors.lastname: Yi
authors.forename: Jiao
authors.initials: J
authors.lastname: Li
authors.forename: Hong-peng
authors.initials: HP
authors.lastname: Han
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D015153:Blotting, Western; D019556:COS Cells; D002645:Chickens; D002522:Chlorocebus aethiops; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D007146:Immunoglobulin lambda-Chains / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D007146:Immunoglobulin lambda-Chains; D011994:Recombinant Proteins