Preparation and Characterisation of Monoclonal Antibodies against Highly Abundant Proteins in Human Parotid Saliva

Objective: To prepare and characterise monoclonal antibodies (mAbs) against highly abundant proteins in human parotid saliva for the depletion of these highly abundant proteins in the future proteomic studies of saliva. Methods: Proteins in human parotid saliva were concentrated by using ultrafiltration and analysed by SDS-PAGE. The protein band between 50-65 kDa was cut, ground and used to immunise BALB/c mice. The mAbs against highly abundant proteins in human parotid saliva were prepared through hybridoma technology and characterised by ELISA and Wetstern blot. Results: Eleven hybridoma cell lines secreting mAbs against highly abundant proteins in human parotid saliva were established, and Western blot assay showed that these antibodies were specific for the highly abundant proteins in human parotid saliva. mAbs against salivary amylase, the most abundant protein in human parotid saliva, were characterised by ELISA and Western blot. Conclusions: mAbs against human highly abundant proteins in parotid saliva were successfully prepared and characterised. The present study provides an approach for using these mAbs for the depletion of highly abundant proteins in human parotid saliva in order to better enrich and visualise lower abundant proteins for the studies of disease-related biomarkers in human saliva proteome. Parole chiave: highly abundant proteins, monoclonal antibody, parotid saliva, proteome