Native leptin protein from mithun (Bos Frontalis) shows serodiagnostic potentiality

With the aim of characterising leptin, the adipose tissue-derived cytokine of mithun (Bos frontalis) at molecular level, the present study was designed to isolate mithun leptin from its adipocytes with subsequent immunobiochemical characterisation of the molecule. In the present study, chemical treatment and ultrasonication technique was adopted for isolating leptin protein from mithun adipose tissue. Purification of the protein was done using affinity column chromatography. The molecular, biophysical and serological characterisation of mithun leptin was carried out by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), Matrix Assisted Laser Desorption Ionization-Time oF Flight MALDI-TOF mass spectroscopy and Western blot. The SDS-PAGE analysis revealed that mithun leptin possesses molecular mass of 16 kDa. Western blot analysis confirmed that the mithun adipocytes possessed the seroreactive leptin protein of 16 kDa. However, MALDI-TOF mass spectroscopy and peptide analysis showed the molecular mass of mithun leptin to be of 17, 214.26 Da 17, 214.26 Da. The seroreactivity of affinitypurified fraction was assessed by plate and dip-stick enzyme-linked immunosorbent assay. The present paper shows the serodiagnostic potential of affinity-purified native leptin protein of mithun for the first time.