Screening and characterization of aptamers of human chronic myeloid leukemia (CML) fusion protein BCR-ABL and its structure analysis / 中国免疫学杂志

Objective:To screen and characterize aptamers against BCR-ABL fusion protein.Methods:A 90bp single stranded DNA( ssDNA) random library was subjected to 13 rounds of selection against BCR-ABL fusion protein by systematic evolution of ligands by expotential enrichment ( SELEX ) method, the selected aptamers were cloned and sequenced.The primary sequences and structure of aptamers were analyzed by Clustal W and DNA Folding Sever and the percentage of the ssDNA pool bound to BCR-ABL core protein were determinated.Results: after 13 rounds selection, the percentage of ssDNA pool bound to BCR-ABL fusion protein increased from 0.3%to 47.1%,the results showed that affinities of the Aptamers were different,the second structure analysis revealed possible stem-loops for binding to BCR-ABL fusion protein,the affinity of aptamer A2 to BCR-ABL fusion protein was highest with Kd values as low as 72 nmol/L.Conclusion:Aptamers against BCR-ABL fusion protein has been identified by SELEX methods from a 90 bp single stranded DNA library.And provide certain reference for the clinical treatment of chronic myelogenous.

SELEX technology and its clinical application / 中华检验医学杂志

SELEX is a newly developed biochemical technique,which filter out high specificity and high affinity ligand for the target molecules through the identification of aptamer combined with the target molecules.The specific aptamer was used in a variety of clinical applications,such as diagnosis of the disease,development of new therapeutic drugs and even directly applied to disease treatment.