IL 6 fold values for all contrasts.

The development of aptamers has been almost exclusively performed based on the SELEX method since their inception. While this method represents a powerful means of harnessing the in vitro evolution of sequences that bind to a given target, there are significant constraints in the design. The most significant constraint has been the reliance on counter selection on off-targets to drive specificity. Counter selection has not been as effective at driving aptamer specificity as the presence of immune tolerance, the capacity of the immune system to remove antibodies that bind to host targets, is for antibody development. This deficiency has constrained the commercial efficacy of aptamers to date. These limitations have been addressed with our design of a novel platform for aptamer identification. This new design is based on what we refer to as a Neomer library with sixteen random nucleotides interspersed with fixed sequences. The fixed sequences are designed to minimize the potential for hybridization, such that secondary structure is driven by the random nucleotides. The use of sixteen random nucleotides reduces the possible library sequences to 4.29 × 109. This enables the application of the same sequences to either the same target or different targets while maintaining a high level of structural diversity. In effect, this introduces the capacity for reproducibility in aptamer selection and an in-silico approach to replicating immune tolerance. We provide here an overview of the new method and a description of the performance of aptamers selected for interleukin 6 developed using this approach.

自适配体（aptamers）诞生以来，其开发几乎完全依托指数富集配体系统进化（SELEX）技术展开。尽管该方法是获取结合特定靶点序列体外进化的有效手段，但其设计存在显著局限。其中最突出的制约因素在于，需依赖针对脱靶靶点的反向筛选以提升适配体特异性。与抗体开发中免疫耐受——即免疫系统清除结合宿主靶点抗体的能力——所发挥的作用相比，反向筛选在提升适配体特异性方面效果欠佳。这一缺陷迄今制约了适配体的商用效能。本研究提出的新型适配体筛选平台已解决了上述局限。该新平台基于我们称之为Neomer文库的设计，由16个随机核苷酸与固定序列交错排布构成。固定序列的设计旨在最大程度降低核酸杂交的可能性，使二级结构仅由随机核苷酸决定。采用16个随机核苷酸可将文库的潜在序列总数缩减至4.29×10⁹。这使得同一套序列既可应用于同一靶点，也可用于不同靶点，同时保持高水平的结构多样性。实际上，这为适配体筛选提供了可重复性的可能，并开创了一种模拟免疫耐受的计算机模拟（in silico）方法。本文概述了该新方法，并介绍了采用此策略筛选得到的针对白细胞介素6（interleukin 6, IL-6）的适配体的性能表现。