title: [The expression, renaturation, purification and preliminary identification of human IgE Cepsilon3-Cepsilon4].
abstract: OBJECTIVE
  To gain recombinant protein Cepsilon3-Cepsilon4 of IgE Fc (E34).
  
  
  METHODS
  We cloned the gene coding human IgE Cepsilon3-Cepsilon4 (E34) and constructed an expression vector pET28a(+)-E34. The target protein was expressed as inclusion body in E.coli BL-21. Following renaturation and purification through a CM sephorose FF column, the soluble protein was acquired, and its binding ability to murine anti-hIgE mAb was identified by Western blot and ELISA.
  
  
  RESULTS
  The cloned E34 gene was sequenced and proved by SDS-PAGE to be the same as reported sequence. SDS-PAGE analysis showed the relative molecular mass of E34 protein obtained was correct as predicted. Western blot and ELISA data revealed that it owned the epitope of binding to murine anti-hIgE mAb.
  
  
  CONCLUSIONS
  The expression vector pET28a(+)-E34 has been successfully constructed and the target protein E34 recognized specifically by murine anti-hIgE mAb is obtained.
authors.affiliation: Department of Molecular Immunology, Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China. bioqcx@eyou.com
authors.forename: Chun-xia
authors.initials: CX
authors.lastname: Qiao
authors.forename: Zhou
authors.initials: Z
authors.lastname: Lin
authors.forename: Mei-ru
authors.initials: MR
authors.lastname: Hu
authors.forename: Rui
authors.initials: R
authors.lastname: Liu
authors.forename: Wei-song
authors.initials: WS
authors.lastname: Qin
authors.forename: Jian-nan
authors.initials: JN
authors.lastname: Feng
authors.forename: Bei-fen
authors.initials: BF
authors.lastname: Shen
mesh_terms: D000911:Antibodies, Monoclonal / Q000378:metabolism; D015153:Blotting, Western; D045744:Cell Line, Tumor; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005786:Gene Expression Regulation; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007073:Immunoglobulin E / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism*; D011485:Protein Binding; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D000911:Antibodies, Monoclonal; D007073:Immunoglobulin E