Aptamer-Based Multiparameter Analysis for Molecular Profiling of Hematological Malignancies

Abstract

The subtypes of hematological malignancies (HM) with minimal molecular profile differences display an extremely heterogeneous clinical course and a discrepant response to certain treatment regimens. Profiling the surface protein markers offers a potent solution for precision diagnosis of HM by differentiating among the subtypes of cancer cells. Herein, we report the use of Cell-SELEX technology to generate a panel of high-affinity aptamer probes that are able to discriminate subtle differences among surface protein profiles between different HM cells. Experimental results show that these aptamers with apparent dissociation constants (K d ) below 10 nM display a unique recognition pattern on different HM subtypes. By combining a machine learning model on the basis of partial least-squares discriminant analysis, 100% accuracy was achieved for the classification of different HM cells. Furthermore, we preliminarily validated the effectiveness of the aptamer-based multiparameter analysis strategy from a clinical perspective by accurately classifying complex clinical samples, thus providing a promising molecular tool for precise HM phenotyping.

Highly Specific Aptamer–Antibody Birecognized Sandwich Module for Ultrasensitive Detection of a Low Molecular Weight Compound

“…Despite these attributes, the false negatives caused by low valence of the antibody and the false positives caused by the complex environment of the sample reduce the credibility of ELISA. , Meanwhile, the enzymatic coloration reaction in the ELISA determines its detection limit at the nanomolar level, rendering ELISA already unable to meet the current demand. , More importantly, the assay is more suitable for the detection of macromolecular weight targets (proteins) rather than low molecular weight compounds (mycotoxins), since two antibodies with large steric hindrance are difficult to simultaneously bind to a small molecule target. Thus, the false negatives are significantly aggravated once the micromolecule is set as the target in ELISA. , Meanwhile, a nucleic-acid-based aptamer, a specific sequence recognizing a target molecule, can directly bind low molecular compounds via the flexible structure . Herein, the aptamer–antibody sandwich assay combined with the high specificity of the antibody and the flexibility of the aptamer structure has been a desirable configuration for detecting low molecular weight compounds.…”