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Identification of Aptamers that Specifically Bind to A1 Antigen by Performing Cell-on Human Erythrocytes

Seyed Mohammad Hasan Hosseini, Mohammad Reza Bassami, Alireza Haghparast, Mojtaba Sankian, Gholamreza Hashemi Tabar

Background: The apply of aptamers as a new generation’s way to probe diagnostic for the detection of target molecules has gained ground. Aptamers can be used as alternatives to diagnostic antibodies for detection of blood groups due to their unique features. This study was aimed to produce DNA diagnostic aptamer detecting the antigen of A1 blood group using the Cell-Selex method. Materials and Methods: DNA aptamer was isolated against A1 RBC antigen after ten stages of Cell-Selex and amplification by an asymmetric polymerase chain reaction. The progress of the stages of selection was evaluated using flow cytometry analysis, which the DNA aptamer isolated from the tenth cycle with an affinity of 70% fluorescent intensity, was selected from four positive colonies followed by determination of the sequences and secondary structures. Results: The aptameric sequence obtained from C4 cloning was calculated with the highest binding affinity to A1 antigen having an apparent dissociation constant (Kd value) of at least 29.5 ± 4.3 Pmol, which was introduced as the selected aptamer-based on ΔG obtained from a colony of C4 equal to –13.13. Conclusion: The aptamer obtained from using Cell-Selex method could be used as an example for the development of diagnostic tools such as biosensors for detecting A1 blood group antigens. [GMJ.2020;9:e1657] 

ABO Blood-Group System; Antibodies; SELEX Aptamer Technique; Flow Cytometry

Procter J, Crawford J, et al. A rapid molecular method (polymerase chain reaction with sequence-specific primers) to genotype for ABO blood group and secretor status and its potential for organ transplants. Tissue Antigens. 1997; 50(5): 475-83.


Hosoi E. Biological and clinical aspects of ABO blood group system. J Med Invest. 2008;55(3-4): 174-82.


Breimer ME, Molne J, et al. Blood group A and B antigen expression in human kidneys correlated to A1/A2/B, Lewis, and secretor status. Transplantation. 2006;82(4): 479-85.


Malomgre W, Neumeister B. Recent and future trends in blood group typing. Anal Bioanal Chem. 2009;393(5): 1443-51.


Mascini M. Aptamers and their applications. Anal Bioanal Chem. 2008;390(4): 987-8.


Jayasena SD. Aptamers: an emerging class of molecules that rival antibodies in diagnostics. Clin Chem. 1999;45(9): 1628-50.


Toh SY, et al. Aptamers as a replacement for antibodies in enzyme-linked immunosorbent assay. Biosens Bioelectron. 2015;64: 392-403.


Sefah K, et al. Development of DNA aptamers using Cell-SELEX. Nat Protoc. 2010;5(6): 1169-85.


Citartan M, et al. Conditions optimized for the preparation of single-stranded DNA (ssDNA) employing lambda exonuclease digestion in generating DNA aptamer. World Journal of Microbiology and Biotechnology. 2011;27(5): 1167-73.

Bai Y, Feng F, et al. Aptamer/thrombin/aptamer-AuNPs sandwich enhanced surface plasmon resonance sensor for the detection of subnanomolar thrombin. Biosens Bioelectron. 2013;47: 265-70.


Citartan M, et al. Asymmetric PCR for good quality ssDNA generation towards DNA aptamer production. Sonklanakarin Journal of Science and Technology. 2012;34(2): 125.

Birch CM, et al. Identification of malaria parasite-infected red blood cell surface aptamers by inertial microfluidic SELEX (I-SELEX). Sci Rep. 2015;5: 11347.

PMid:26126714 PMCid:PMC4486934

Nuo Duan, et al. Selection, identification and application of a DNA aptamer against Listeria monocytogenes. Food Control . 2013;33: 239-243.

Dwivedi HP, et al. Selection of DNA aptamers for capture and detection of Salmonella Typhimurium using a whole-cell SELEX approach in conjunction with cell sorting. Appl Microbiol Biotechnol. 2013;97(8): 3677-86.


Kazunori Ikebukuro, et al. Selection of DNA aptamer against prostate specific antigen using a genetic algorithm and application to sensing. Biosensors and Bioelectronics. 2010;26: 1386-1391.


Jenkins PV, O'Donnell JS. ABO blood group determines plasma von Willebrand factor levels: a biologic function after all? Transfusion. 2006;46(10): 1836-44.


Zuker M. Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Res. 2003;31(13): 3406-15.

PMid:12824337 PMCid:PMC169194

Duan N, et al. In vitro selection of a DNA aptamer targeted against Shigella dysenteriae. J Microbiol Methods. 2013;94(3): 170-4.


Sun MQ, et al. DNA aptamer selection in vitro for determining ketamine by FluMag-SELEX. Fa Yi Xue Za Zhi. 2014;30(5): 346-9.

Vahidi H, et al. Challenges to Design and Develop of DNA Aptamers for Protein Targets. II. Development of the Aptameric Affinity Ligands Specific to Human Plasma Coagulation Factor VIII Using SEC-SELEX. Iran J Pharm Res. 2017;16(2):737-44.

Nimjee SM, et al. Aptamers: an emerging class of therapeutics. Annu Rev Med. 2005;56: 555-83.


Meirinho SG, et al. Development of an electrochemical RNA-aptasensor to detect human osteopontin. Biosens Bioelectron. 2015;71: 332-41.


D'Andrea D, et al. ABO Blood Group and Rhesus Factor Are Not Associated with Outcomes After Radical Cystectomy for Non-metastatic Urothelial Carcinoma of the Bladder. Anticancer Res. 2017;37(10): 5747-53.

Ni X, et al. Nucleic acid aptamers: clinical applications and promising new horizons. Curr Med Chem. 2011;18(27): 4206-14.

PMid:21838685 PMCid:PMC3260938


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