Abstract

ISOLATION AND PURIFICATION OF ANTI-H LECTIN FROM MOMORDICA CHARANTIA SEEDS AND ITS DIAGNOSTIC USE FOR DIFFERENTIATING BOMBAY PHENOTYPE (Oh) BLOOD GROUP FROM O BLOOD GROUP

Rohan Sharadanand Phatak*, Ajit Pandurang Purohit, Pushpa Prakash Durgawale, Anup Subhash Hendre, Sachin Eknath Potawale

ABSTRACT

Background: In 1952, Bhende, Bhatia, and Deshpande identified a new blood group called Bombay Phenotype blood group Oh which is characterized by the absence of A, B and H antigens on the red blood cells. Therefore it is extremely difficult for finding of compatible Oh red blood cells for blood transfusion as Bombay blood group cells do not agglutinate even from serum of other Bombay blood group people since they lack the H antigen on their red blood cells. The effort was made to provide anti-H lectin extracted from Momordica charantia seeds in a simple isolation process, less time-consuming, economical purification method. Material and Methods: In this study, Momordica charantia seed powder was homogenized in phosphate buffered saline (PBS); filtrate was centrifuged and supernatant layer was precipitated with ammonium sulphate crystals to get lectin. Lectin was isolated and purified by method of formalinised erythrocytes affinity chromatography. Galactose was used to elute anti-H specific lectin from formalinised erythrocytes affinity chromatography. Haemagglutinating activity of Momordica charantia lectin was tested among all blood groups such as A, B, AB, O and Bombay Phenotype blood groups. Thermal stability, SDS-PAGE and PAGE electrophoresis of Momordica charantia lectin were carried out. Ulex europaeus lectin was used as standard anti-H specific lectin for detecting Bombay Phenotype blood group. Results: Formalinised erythrocytes affinity chromatography is better method to isolate anti-H specific lectin than acetone precipitation method. By comparing with Ulex europaeus lectin, Momordica charantia lectin has easily available, higher percentage yield, stronger haemagglutinating activity, better thermal stability and has low cost to manufacture. Therefore, Momordica charantia lectin has more commercial value than that of Ulex europaeus lectin. Conclusion: Since Ulex europaeus lectin is very expensive only few blood banks use Ulex europaeus lectin routinely to differentiate O blood group from Bombay blood group. If a cheaper and better quality product like Momordica charantia lectin is made available, it will be routinely and regularly used in all the blood banks by preventing hemolysis risks. Therefore, Momordica charantia lectin is commercially more viable product.

Keywords: Bombay Phenotype Blood Group, Lectin, Momordica charantia, Ulex europaeus.