[Preparation and activity detection of chicken egg yolk IgY antibody against human papillomavirus 16 type L1 main capsid protein].
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.
Yang
Jun
J
Department of Pathology, Second Hospital Affiliated to Xi'an Jiaotong University College of Medicine, Xi'an 710004, China. yangjundr@yahoo.com
Zhang
Ming-juan
MJ
Qiang
Lei
L
Su
Bao-shan
BS
Wang
Yi-li
YL
Si
Lü-sheng
LS
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
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