Ethics code: IR.TMI.REC.1395.016
Abstract: (21 Views)
gen derived from a synthetic peptide specific to the D antigen. Ultimately, the potential of these antibodies to agglutinate human red blood cells was investigated.
Materials and Methods
In this experimental study, antibodies against RhD were generated in rabbits. Rabbits 1 and 2 were immunized with the complete RhD antigen, which was purified from the membranes of red blood cells by affinity chromatography. Concurrently, Rabbit 3, was immunized with an RhD antigen dipeptide, which was prepared from a selected peptide of RhD using the O-PDM method. IgG molecules were purified from the antisera produced in rabbits. The characteristics of both the purified RhD antigen and the antisera were evaluated using the ELISA method. Additionally, the functional activity of antisera was assessed through the agglutination of human red blood cells.
Results
The characteristic of the purified RhD antigen was demonstrated by ELISA assay using commercial anti-RhD antibody. Antibody formation in the sera of rabbits was also confirmed using the ELISA method. The isolated IgG polyclonal rabbit antibodies against RhD showed agglutination of O+ red blood cells in the presence of a secondary antibody (n=3). Subsequently, the specificity of the produced antisera was similar between rabbits immunized with the purified antigen and those immunized with the dimerized peptide. The antibodies were only able to agglutinate red blood cells through a two-step method. The synthetic peptide related to the amino terminus of RhD antigen, in its dimeric form, exhibited immunogenic characteristic comparable to those of the complete antigen.
Conclusions
These findings indicate that the appropriate dimeric form of a peptide can serve as a substitute for the conjugated peptide and a carrier protein for achieving successful immunization.
Materials and Methods
In this experimental study, antibodies against RhD were generated in rabbits. Rabbits 1 and 2 were immunized with the complete RhD antigen, which was purified from the membranes of red blood cells by affinity chromatography. Concurrently, Rabbit 3, was immunized with an RhD antigen dipeptide, which was prepared from a selected peptide of RhD using the O-PDM method. IgG molecules were purified from the antisera produced in rabbits. The characteristics of both the purified RhD antigen and the antisera were evaluated using the ELISA method. Additionally, the functional activity of antisera was assessed through the agglutination of human red blood cells.
Results
The characteristic of the purified RhD antigen was demonstrated by ELISA assay using commercial anti-RhD antibody. Antibody formation in the sera of rabbits was also confirmed using the ELISA method. The isolated IgG polyclonal rabbit antibodies against RhD showed agglutination of O+ red blood cells in the presence of a secondary antibody (n=3). Subsequently, the specificity of the produced antisera was similar between rabbits immunized with the purified antigen and those immunized with the dimerized peptide. The antibodies were only able to agglutinate red blood cells through a two-step method. The synthetic peptide related to the amino terminus of RhD antigen, in its dimeric form, exhibited immunogenic characteristic comparable to those of the complete antigen.
Conclusions
These findings indicate that the appropriate dimeric form of a peptide can serve as a substitute for the conjugated peptide and a carrier protein for achieving successful immunization.
Send email to the article author
| Rights and permissions | |
 |
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
