Abstract
Cirrhosis of liver is an end stage of chronic liver insults from varied etiologies which leads to impaired liver functions. Proteins expressed from liver and enters into circulation reflects degree of liver dysfunction. Serpins (Serine protease inhibitors) are class of plasma proteins expressed from liver; SERPINA4/Kallistatin is a multifunctional serpin clade A protein expressed from liver and concentration in serum is the reflection of extent of liver dysfunction. The present study aimed to compare cross reactivity of serpins for polyclonal and monospecific antibodies in both cirrhotic liver and healthy subjects. Blood samples were collected from 20 subjects (10 cirrhotic liver, 10 healthy) from R. L. Jalappa Hospital and Research Centre, Kolar, Karnataka, India. Separation of proteins was carried out by SDS-PAGE. Cross reactivity study was analyzed using western blot. Proteins present in cirrhotic liver and healthy subject’s serum were separated by SDS PAGE. There was no band detection on both (cirrhotic liver and healthy) PVDF (polyvinylidene diflouride) membranes with polyclonal antibodies. However, a significant band was observed with protein of interest in healthy PVDF membrane with monospecific antibodies. There was no band in cirrhotic liver PVDF membrane even with monospecific antibodies. Comparative cross reactivity analysis of serpins in quantification of SERPINA4/Kallistatin in the present study demonstrated that there will not be any immunological cross reactivity between serpins and SERPINA4/Kallistatin due to the absence of identical epitope in cirrhotic liver and healthy subjects.
Highlights
Cirrhosis of liver is reversible natural wound healing response which results in the formation of abnormal continuation of connective tissue production and deposition and regenerative nodular formation in response to chronic liver injury [1, 2]
Individuals diagnosed with cirrhosis of liver caused by alcoholic liver disease (ALD) and non-alcoholic fatty liver diseases (NAFLD) based on clinical history and symptoms viz., ascites, encephalopathy, jaundice and altered biochemical parameters were included in the study
Western blot analysis allowed identification of cross reactivity of serpins with SERPINA4/Kallistatin in diseased and healthy samples by using polyclonal antibodies specific for SERPINA4/Kallistatin followed by secondary antibodies conjugated with HRP
Summary
Cirrhosis of liver is reversible natural wound healing response which results in the formation of abnormal continuation of connective tissue production and deposition and regenerative nodular formation in response to chronic liver injury [1, 2]. Reactive centre loop (RCL) of these proteins in secondary structure interacts with active site of proteases and inhibits their action. They undergo conformational changes in their structure which is crucial for their activity. Apart from inhibitory action, they acts as transporters for hormones, plays an important role in coagulation and blood pressure regulation [5]
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