[Preparation and identification of type specific and conformation dependent HPV16 L1 protein monoclonal antibody.].
OBJECTIVE
To generate type specific and conformation dependent monoclonal antibodies against human papillomavirus 16 major capsid L1 protein (HPV16 L1).


METHODS
HPV16 L1 protein was expressed by modified insect-baculovirus expression system and purified by ProBond(TM); purification system in nature conditions. BALB/c mice were immunized with the purified HPV16 L1 protein. Hybridoma technique was used to prepare the hybridoma cell lines. Positive colonies were screened by ELISA. The non-denaturing electrophoresis and Western blot methods were used for detecting the conformation dependent antibodies. Immunocytochemistry was used to confirm the type specific of the antibodies. The sub-class, isotype and titer were also identified respectively.


RESULTS
One hybridoma cell line named 2E3 was obtained which can constantly secret type-specific and conformation-dependent HPV16 L1 mAb. The titers of the antibody are 1:800 in the cell culture medium and 1:12 800 in ascetic fluid. The sub-class and isotype were IgG1kappa. The antibody could selectively recognize non-denaturing HPV16 L1 protein and there was no cross-reaction with HPV18, HPV58 and HPV11 L1 protein.


CONCLUSIONS
One cell line which secretes type specific and conformation dependent HPV16 L1 monoclonal antibody is successfully generated. This monoclonal antibody can be used in study HPV16 L1 protein epitope.
Yan
Xiao-Fei
XF
Center for Cancer Research, the First Affilicated Hospital, School of Life Science and Technology , Xi'an Jiaotong University, Xi'an 710061, China.
Xu
Yang
Y
Zheng
Jin
J
Lai
Bao-Chang
BC
Wang
Yi-Li
YL
D000818:Animals; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D036022:Capsid Proteins / Q000276:immunology; D052162:Human papillomavirus 16; D006801:Humans; D008807:Mice, Inbred BALB C; D009856:Oncogene Proteins, Viral / Q000276:immunology; D027383:Papillomaviridae
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D036022:Capsid Proteins; D009856:Oncogene Proteins, Viral