Volume 20, Issue 1 (Spring 2023)
Sci J Iran Blood Transfus Organ 2023, 20(1): 28-33 |
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Abedini K, Yari F, Milani S. Investigating the solubilization conditions of packed red blood cells to obtain Duffy a antigen in the laboratory. Sci J Iran Blood Transfus Organ 2023; 20 (1) :28-33
URL: http://bloodjournal.ir/article-1-1464-en.html
URL: http://bloodjournal.ir/article-1-1464-en.html
Abstract: (1078 Views)
Abstract
Background and Objectives
One of the antigens of the blood group is Duffy, which includes different subtypes. This system is of clinical importance in terms of blood transfusion, so accurate identification of this antigen is very important. In this study, the process of solubilization of red blood RBC membrane and purification of the Duffy antigen, were investigated.
Materials and Methods
In this experimental study, packed RBCs (O- and Fya+) were obtained from the blood donation center of Tehran province. The membranes of RBCs were solubilized using Nonidet P-40 detergent. In the next step, Fya antigen was purified by affinity chromatography method using a specific antibody against this antigen. The specificity of the purified protein was tested by ELISA technique.
Results
The Fya+ characteristic of red blood cells of the RBC bag was confirmed. In addition, the specificity of the antigen was confirmed by ELISA method. The absorbance levels of the purified antigen (OD450nm) in ELISA method were 1.056 ± 0.137 and 0.197 ± 0.016 by employing the Fya and Fyb antibodies, respectively.
Conclusions
The present research showed that red blood cell membrane solubilization can be done to purify Fy antigen and this antigen may be used in various fields including immunization and antibody production.
Background and Objectives
One of the antigens of the blood group is Duffy, which includes different subtypes. This system is of clinical importance in terms of blood transfusion, so accurate identification of this antigen is very important. In this study, the process of solubilization of red blood RBC membrane and purification of the Duffy antigen, were investigated.
Materials and Methods
In this experimental study, packed RBCs (O- and Fya+) were obtained from the blood donation center of Tehran province. The membranes of RBCs were solubilized using Nonidet P-40 detergent. In the next step, Fya antigen was purified by affinity chromatography method using a specific antibody against this antigen. The specificity of the purified protein was tested by ELISA technique.
Results
The Fya+ characteristic of red blood cells of the RBC bag was confirmed. In addition, the specificity of the antigen was confirmed by ELISA method. The absorbance levels of the purified antigen (OD450nm) in ELISA method were 1.056 ± 0.137 and 0.197 ± 0.016 by employing the Fya and Fyb antibodies, respectively.
Conclusions
The present research showed that red blood cell membrane solubilization can be done to purify Fy antigen and this antigen may be used in various fields including immunization and antibody production.
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