title: [Preparation and activity detection of chicken egg yolk IgY antibody against human papillomavirus 16 type L1 main capsid protein].
abstract: OBJECTIVE
  To prepare highly specific chicken egg yolk IgY antibody against human papillomavirus 16 type L1 main capsid protein (HPV16L1) for detection of HPV16L1.
  
  
  METHODS
  Purified HPV16L1 protein was used to immunize the hens, from which the eggs were collected since one week after the first immunization. The egg yolk was separated and the IgY antibody purified by PEG-6000 method. The bioactivity of the antibody was tested using enzyme-linked immunosorbent assay (ELISA). Immunohistochemistry was performed to detect the HPV16L1 in the CHO cells transfected with the recombinant pcDNA-EGFP-HPV16L1 plasmid (containing EGFP-HPV16L1 fusion gene) for assessing the specific affinity of IgY to HPV16L1.
  
  
  RESULTS
  After 3 immunizations of the hens, the titer of the purified IgY antibody against HPV16L1 from the egg yolk reached 1:10240. The IgY bound specifically to the EGFP-HPV16L1 protein expressed in the transfected CHO cells.
  
  
  CONCLUSIONS
  High titer IgY can be prepared by immunization of the hens with HPV16L1 protein, and the prepared IgY can be used for HPV16L1 detection at the cellular level.
authors: Department of Pathology, Second Hospital Affiliated to Xi'an Jiaotong University College of Medicine, Xi'an 710004, China. yangjundr@yahoo.com
authors: Jun
authors: J
authors: Yang
authors: Ming-juan
authors: MJ
authors: Zhang
authors: Lei
authors: L
authors: Qiang
authors: Bao-shan
authors: BS
authors: Su
authors: Yi-li
authors: YL
authors: Wang
authors: Lü-sheng
authors: LS
authors: Si
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis / Q000276:immunology* / Q000302:isolation & purification; D000918:Antibody Specificity / Q000276:immunology; D016466:CHO Cells; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D002645:Chickens; D006224:Cricetinae; D003412:Cricetulus; D004797:Enzyme-Linked Immunosorbent Assay; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007114:Immunization / Q000379:methods; D007136:Immunoglobulins / Q000276:immunology*; D007150:Immunohistochemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D009856:Oncogene Proteins, Viral / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D014162:Transfection
chemical_list: D000914:Antibodies, Viral; D036022:Capsid Proteins; C016512:IgY; D007136:Immunoglobulins; D009856:Oncogene Proteins, Viral; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; C063317:L1 protein, Human papillomavirus type 16