Preparation and activity detection of chicken egg yolk IgY antibody against human papillomavirus 16 type L1 main capsid protein.
- Author:
Jun YANG
1
;
Ming-juan ZHANG
;
Lei QIANG
;
Bao-shan SU
;
Yi-li WANG
;
Lü-sheng SI
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Antibodies, Viral; analysis; immunology; isolation & purification; Antibody Specificity; immunology; CHO Cells; Capsid Proteins; genetics; immunology; metabolism; Chickens; Cricetinae; Cricetulus; Enzyme-Linked Immunosorbent Assay; Green Fluorescent Proteins; genetics; immunology; metabolism; Humans; Immunization; methods; Immunoglobulins; immunology; Immunohistochemistry; Mice; Mice, Inbred C57BL; Oncogene Proteins, Viral; genetics; immunology; metabolism; Recombinant Fusion Proteins; genetics; immunology; metabolism; Transfection
- From: Journal of Southern Medical University 2008;28(3):324-327
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo prepare highly specific chicken egg yolk IgY antibody against human papillomavirus 16 type L1 main capsid protein (HPV16L1) for detection of HPV16L1.
METHODSPurified 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.
RESULTSAfter 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.
CONCLUSIONHigh 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.
