title: [Synthesis and identification of artificial complete antigen 25-hydroxyvitamin D(3);].
abstract: OBJECTIVE
  To synthesize the 25-hydroxyvitamin D(3); artificial complete antigen and to prepare the specific antibody against 25-hydroxyvitamin D(3);.
  
  
  METHODS
  The active group carboxyl was introduced into 25-hydroxyvitamin D(3); and formed 25-hydroxyvitamin D(3);-hemisuccinate which possessed the structure of the hapten by chemical modification. The EDC method was applied to conjugate 25-hydroxyvitamin D(3);-hemisuccinate to bovine serum albumin as an artificial immunogen. The coating antigen 25-hydroxyvitamin D(3);-hemisuccinate-OVA was obtained in the same way. Ultraviolet,  SDS-PAGE and MALDI-TOF were used to identify 25-hydroxyvitamin D(3);-hemisuccinate-BSA.
  
  
  RESULTS
  BALB/c mice were immunized with 25-hydroxyvitamin D(3);-hemisuccinate-BSA to generate the polyclonal antibody of the 25-hydroxyvitamin D(3); worth high titer and the immunogen,  25-hydroxyvitamin D(3);-hemisuccinate-BSA,  was successfully prepared with coupling ratio (12±0.16):1(N=3) coupling.
  
  
  CONCLUSIONS
  The high titer and good sensitivity of anti-25-hydroxyvitamin D(3); antibody are produced in sera immunized BALB/c mice,  which made it possible to develop a clinical diagnostics for illness.
authors.affiliation: Institute For Biomedical Research, Xiamen University, Xiamen, China.
authors.forename: Wei-wei
authors.initials: WW
authors.lastname: Wan
authors.forename: Huan-jing
authors.initials: HJ
authors.lastname: Zhang
authors.forename: Hui-ming
authors.initials: HM
authors.lastname: Ye
authors.forename: Li-qin
authors.initials: LQ
authors.lastname: Wan
authors.forename: Liang-liang
authors.initials: LL
authors.lastname: Cai
authors.forename: Li-mou
authors.initials: LM
authors.lastname: Zheng
authors.forename: Chi-meng
authors.initials: CM
authors.lastname: Zeng
mesh_terms: D000818:Animals; D000906:Antibodies / Q000097:blood*; D000918:Antibody Specificity; D000941:Antigens / Q000378:metabolism; D002112:Calcifediol / Q000138:chemical synthesis* / Q000737:chemistry* / Q000276:immunology; D002417:Cattle; D002762:Cholecalciferol / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry; D005260:Female; D007114:Immunization / Q000379:methods; D051379:Mice; D008807:Mice, Inbred BALB C; D010047:Ovalbumin / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D013386:Succinates / Q000138:chemical synthesis* / Q000737:chemistry
chemical_list: C108995:25-hydroxyvitamin D3 3-hemisuccinate; D000906:Antibodies; D000941:Antigens; D013386:Succinates; D002762:Cholecalciferol; D012710:Serum Albumin, Bovine; D010047:Ovalbumin; D002112:Calcifediol