title: [Preparation and activity detection of monoclonal antibody against anti-CD3 ScFv].
abstract: OBJECTIVE
  To prepare monoclonal antibody (McAb) against anti-CD3 ScFv for purifying and detecting serum anti-CD3 antibody concentration.
  
  
  METHODS
  McAb against anti-CD3 ScFv was prepared by hybridoma technique and used to prepare affinity chromatography column, which was used to purify anti-CD3 ScFv and Diabody [CD3 x Pgp] without E-tag. The binding activities of anti-CD3 ScFv, Diabody [CD3 x Pgp] without E-tag, and Diabody [CD3 x Pgp] purified by anti-CD3 affinity chromatography column or anti-E-tag affinity chromatography column against K562/A02 cell and Jurket cells were detected by fluorescence activated cell sorting (FACS) method. ELISA was used to identify the specificity of the McAb.
  
  
  RESULTS
  McAb against anti-CD3 ScFv specifically detected serum anti-CD3 ScFv without reacting with sera. The anti-CD3 ScFv purified by anti-CD3 affinity chromatography column and purified by anti-E-tag affinity chromatography column had the same specific binding activity with Jurkat cells. The positive binding rates of Diabody [CD3 x Pgp] without E-tag to K562/A02 and Jurkat cells were 89.87% and 83.95%, respectively. In the competitive binding experiments with K562/A02 and Jurkat cells, the binding rates of Diabody [CD3 x Pgp] without E-tag decreased to 56.30% and 43.78%, respectively.
  
  
  CONCLUSIONS
  The McAb against anti-CD3 ScFv prepared in our lab can be used to purify and detect serum anti-CD3 antibody concentration.
authors.affiliation: State Key Laboratory of Experimental Hematology, Institute of Hematology, CAMS and PUMC, Tianjin 300020, China.
authors.forename: Xiao-Feng
authors.initials: XF
authors.lastname: Shao
authors.forename: Ying-Dai
authors.initials: YD
authors.lastname: Gao
authors.forename: Juan-Ni
authors.initials: JN
authors.lastname: Liu
authors.forename: Jin-Hong
authors.initials: JH
authors.lastname: Wang
authors.forename: Yuan-Fu
authors.initials: YF
authors.lastname: Xu
authors.forename: Dong-Mei
authors.initials: DM
authors.lastname: Fan
authors.forename: Chun-Zheng
authors.initials: CZ
authors.lastname: Yang
authors.forename: Dong-Sheng
authors.initials: DS
authors.lastname: Xiong
mesh_terms: D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology* / Q000302:isolation & purification; D017252:CD3 Complex / Q000276:immunology*; D002460:Cell Line; D002846:Chromatography, Affinity; D006801:Humans; D006825:Hybridomas / Q000378:metabolism; D019169:Jurkat Cells; D020014:K562 Cells
chemical_list: D000911:Antibodies, Monoclonal; D017252:CD3 Complex