Lafuente-González Elsa, Guadaño-Sánchez Miriam, Urriza-Arsuaga Idoia, Urraca Javier Lucas
Independent Researcher, E-28040 Madrid, Spain.
Chemical Optosensors and Applied Photochemistry Group (GSOLFA), Department of Analytical Chemistry, Facultad de Química, Universidad Complutense de Madrid, E-28040 Madrid, Spain.
Int J Mol Sci. 2023 Feb 9;24(4):3453. doi: 10.3390/ijms24043453.
FLAG tag (DYKDDDDK) is a small epitope peptide employed for the purification of recombinant proteins such as immunoglobulins, cytokines, and gene regulatory proteins. It provides superior purity and recoveries of fused target proteins when compared to the commonly used His-tag. Nevertheless, the immunoaffinity-based adsorbents required for their isolation are far more expensive than the ligand-based affinity resin used in combination with the His-tag. In order to overcome this limitation we report herein the development of molecularly imprinted polymers (MIPs) selective to the FLAG tag. The polymers were prepared by the epitope imprinting approach using a four amino acids peptide, DYKD, including part of the FLAG sequence as template molecule. Different kinds of magnetic polymers were synthesised in aqueous and organic media also using different sizes of magnetite core nanoparticles. The synthesised polymers were used as solid phase extraction materials with excellent recoveries and high specificity for both peptides. The magnetic properties of the polymers confer a new, effective, simple, and fast method in the purification using FLAG tag.
FLAG标签（DYKDDDDK）是一种小表位肽，用于纯化重组蛋白，如免疫球蛋白、细胞因子和基因调节蛋白。与常用的His标签相比，它能提供更高纯度的融合靶蛋白，回收率也更高。然而，分离它们所需的基于免疫亲和的吸附剂比与His标签结合使用的基于配体的亲和树脂要昂贵得多。为了克服这一限制，我们在此报告了对FLAG标签具有选择性的分子印迹聚合物（MIP）的开发。这些聚合物是通过表位印迹方法制备的，使用一种四氨基酸肽DYKD，其包含FLAG序列的一部分作为模板分子。还使用不同尺寸的磁铁矿核心纳米颗粒在水性和有机介质中合成了不同种类的磁性聚合物。合成的聚合物用作固相萃取材料，对两种肽都具有优异的回收率和高特异性。聚合物的磁性为使用FLAG标签的纯化提供了一种新的、有效、简单且快速的方法。
