Preparation and Purification of the Monoclonal Antibody for Anti-human Lung Squamous Carcinoma

Objective To subclone C5 hybridoma cells and purify the monoclonal antibody(McAb) by the immunoaffinity chromatography so as to provide with purified McAb the future research,the early diagnosis of lung cancers and targeted therapy and to study the lung cancer associated with antigen and the antigen associated with vaccine.Method C5 hybridoma cells were thawed from liquid nitrogen.The culture of hybridoma cells was subcloned for 3 times,and the culture's supernatant from each clone was tested by ELISA assay.Positive clones underwent culture expansion and the McAb was purified from its supernatant using ProteinaA-SepharoseCL-4B column affinity chromatography procedure.The specificity of the McAb protein expression was measured using SDS-PAGE and Western-Blot assay.Results the McAb was purified from C5 lung cancer hybridoma cell line using affinity chromatography technique.SDS-PAGE and Western Blot assay of the McAb showed one single band was specifically against the second antibody and the molecular weight of which was about 75KDa.Conclusion The McAb against lung cancer cells can be successfully purified from C5 hybridoma cells by limited dilution and affinity chromatography using ProteinaA-SepharoseCL-4B column.This McAb may play an important role in the early diagnosis and targeted therapy of human lung cancer and may contribute to the development of tumor vaccine.