Ruff Laura E, Sapre Ajay A, Plaut Justin S, De Maere Elisabeth, Mortier Charlotte, Nguyen Valerie, Separa Kevin, Vandenbogaerde Sofie, Vandewalle Laura, Esener Sadik C, Messmer Bradley T
UCSD Moores Cancer Center, University of California San Diego, La Jolla, CA 92093, USA
Department of Bioengineering, University of California San Diego, La Jolla, CA 92093, USA.
Nucleic Acids Res. 2016 Jun 2;44(10):e96. doi: 10.1093/nar/gkw136. Epub 2016 Mar 11.
High affinity and specificity are considered essential for affinity reagents and molecularly-targeted therapeutics, such as monoclonal antibodies. However, life's own molecular and cellular machinery consists of lower affinity, highly multivalent interactions that are metastable, but easily reversible or displaceable. With this inspiration, we have developed a DNA-based reagent platform that uses massive avidity to achieve stable, but reversible specific recognition of polyvalent targets. We have previously selected these DNA reagents, termed DeNAno, against various cells and now we demonstrate that DeNAno specific for protein targets can also be selected. DeNAno were selected against streptavidin-, rituximab- and bevacizumab-coated beads. Binding was stable for weeks and unaffected by the presence of soluble target proteins, yet readily competed by natural or synthetic ligands of the target proteins. Thus DeNAno particles are a novel biomolecular recognition agent whose orthogonal use of avidity over affinity results in uniquely stable yet reversible binding interactions.
高亲和力和特异性被认为是亲和试剂和分子靶向治疗药物（如单克隆抗体）所必需的。然而，生命自身的分子和细胞机制是由低亲和力、高度多价的相互作用组成的，这些相互作用是亚稳态的，但很容易逆转或被取代。受此启发，我们开发了一种基于DNA的试剂平台，该平台利用大量亲和力来实现对多价靶标的稳定但可逆的特异性识别。我们之前已针对各种细胞筛选出了这些被称为DeNAno的DNA试剂，现在我们证明，针对蛋白质靶标的DeNAno也可以被筛选出来。针对链霉亲和素、利妥昔单抗和贝伐单抗包被的珠子筛选出了DeNAno。结合在数周内保持稳定，不受可溶性靶蛋白存在的影响，但很容易被靶蛋白的天然或合成配体竞争。因此，DeNAno颗粒是一种新型的生物分子识别剂，其对亲和力的正交利用导致了独特的稳定但可逆的结合相互作用。
