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Autoimmune haemolytic anaemia is a condition in which autoantibodies present against self erythrocytes which leads to severe haemolysis and loss of haemoglobin. In the present study we compared the electrophoretic band patterns between control and autoimmune haemolytic anaemia and non autoimmune iron deficiency anaemia erythrocyte lysate samples. We observed the presence of a band which was specific to autoimmune haemolytic erythrocyte lysates in the region of 70 Kd which was later identified as albumin and was distinctly absent in control as well as iron deficiency non autoimmune haemolytic samples. In addition, we have noticed that the catalase in the region of 60 Kd was found to be decreased in its band intensity in haemolytic anaemia samples compared to control. The presence of both albumin and catalase in the respective positions in the electrophoretic gel was confirmed by in gel trypsin digestion followed by mass spectrometry. The other erythrocyte proteins visible in the region of 25 Kd were found to have an overall decrease in band intensity in haemolytic anaemia samples compared to control. The specific presence of albumin in autoimmune haemolytic erythrocyte lysate is the main highlight of the work which was not reported till date and further studies could reveal the exact role of albumin in autoimmune haemolytic conditions. Key words: Autoimmune haemolytic anaemia, erythrocytes, electrophoresis, albumin, catalase

Title: Electrophoretic Protein Analysis of Red blood cell Membrane Proteins upon Ionic and Non-ionic detergent lysis Authors & Affiliation Mathew John1, Nimmy Mohandas2, Sinju R3 1, 2 JCMR Jubilee Mission Medical College & Research Institute, Thrissur - 680 005, Kerala, India 3 Department of Biochemistry, Jubilee Mission Medical College & Research Institute, Thrissur - 680 005, Kerala, India mathewjohn@jmmc.ac.in, nimmymohandas6@gmail.com, sinjurajan7@gmail.com Abstract: Red blood cells (RBC) are anucleated atypical cellular architecture with a plasma membrane envelope where the glycoproteins are oriented along the lipid bilayer. RBC proteins are classified as peripheral and integral proteins based on its location along plasma membrane. The present study explored the extraction efficiency of various detergents such as Triton X100, Sodium dodecyl sulphate (SDS), Cetypyridinium chloride (CPC), alone and its combinations along with protease inhibitor cocktail for RBC membrane proteins analyzed in an elctrophoretogram. The study showed that RBC protein extraction was much better in combination of Triton X100 and SDS as evident by increased number of protein bands in coomassie brilliant blue R stained SDS gel, compared to CPC combined Triton X100 lysis buffer. The study hence showed that RBC proteins can be extracted with combination of non-ionic detergent and anionic detergent. Key words: Red blood cells, Triton X100, Sodium dodecyl sulphate, Cetyl pyridinium chloride URL:http://www.ijbst.org/Home/papers-published/ijbst-2018-volume-11-issue-6