Preparation and identification of type specific and conformation dependent HPV16 L1 protein monoclonal antibody

AIM: 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.

CONCLUSION: 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.