Isolation, cloning, expression and purification of recombinant RhD antigen from cord blood

Rh [Rhesus] is a highly complex blood group system in man deeply rooted in transfusion medicine. Isolation of RhD from cord blood, cloning and expression of recombinant RhD antigen in bacterial expression system was the aim of this study. Total RNAs were extracted from cord blood [O[+]]. The quality of RNA was determined by electrophoresis. In order to obtain coding sequence of RhD antigen cDNA was synthesized and Rh D gene was amplified by RT-PCR. The isolated RhD gene was cloned to pUCIS vector and transformed to DH5alpha. The confirmed construct was sub cloned into expression vector, pBADgIII/A, and expressed in Top 10 E.coli. The expressed protein was characterized by SDS-PAGE and western blot analysis. Antigenicity of the expressed protein was assessed by ELISA using commercially available human anti-RhD polyclonal antibody with peroxidase conjugated goat anti-human IgG, IgM, IgA as secondary antibody. RhD gene was successfully cloned and expressed. The expected size of recombinant RhD protein was detected in SDS-PAGE, and confirmed by dot and western blot analysis. RhD antibody reacted with recombinant RhD antigen as well as with RhD polypeptide extracted from RBCs membrane. The recombinant RhD may be helpful to further investigate the molecular basis of RhD protein and could be applicable for production anti- D antibody in an animal model

Satisfaction evaluation of blood donors of Iranian Blood Transfusion Organization in 2003

Satisfaction of blood donors plays a critical role in providing safe and adequate blood supply for patients. This study aimed to evaluate the level of satisfaction of blood donors in Iranian Blood Transfusion Organization [IBTO]. This was a cross-sectional study with the involvement of 2508 blood donors selected in proportion to the average distribution of blood donors in different provinces of Iran. The random sampling method was used. Questionnaires were directly handed over to blood donors. There was a positive relationship between behavior of physicians or other care staff with satisfaction level of blood donors. Physical conditions of donation sites play an important role in satisfaction level of blood donors. There was a negative correlation between education level and satisfaction of blood donors; the higher the education level, the less the rate of satisfaction. There was not any significant difference in satisfaction of blood donors in terms of sex. Satisfaction rate of blood donors was higher among those referring to blood drives than those to blood centers. Satisfaction rate among repeat blood donors was more than twice that among first-time blood donors [PVO.04]. The highest satisfaction value is attributed to the behavior of the personnel involved in phlebotomy and physicians. The lowest satisfaction value pertained to accessibility of blood donor centers. Overall satisfaction rate was more than 50% showing a figure, higher than our previous estimation

[Isolation and detection of RhD antigen from red blood cell [RBC] membrane by immune- precipitation method]

Rh [Rhesus] is a highly complex blood group system in man which plays an important role in transfusion medicine. The aim of this study was the isolation of RhD protein from the membrane of RBCs. In this experimental study immunoprecipitation method with human anti-RhD polyclonal antibody was utilized for the isolation of RhD antigen from Rh[+] human blood samples Proteins of RBCs were characterized by SDS-PAGE and Western blot analysis. Antigenicity of the RhD protein was assessed by ELISA using commercially available human anti-RhD polyclonal antibody with peroxidase conjugated goat anti-human as a secondary antibody. The results show that RhD protein has successfully been isolated by immunoprecipitation method. The expected size of RhD protein was confirmed by Western blot analysis. RhD antibody reacted with RhD antigen prepared from ghost with polyclonal antibody in ELISA, but no reaction was observed in Western blot analysis with monoclonal antibody: It is necessary to mention that this is the primary report of relative purification of RhD and further studies are recommended. The RhD may be helpful to further investigate the molecular basis of RhD protein and could be applicable for production of anti-D antibody in an animal model