A molecularly imprinted polymer as an antibody mimic with affinity for lysine acetylated peptides

Abstract

Lysine acetylation is a widespread protein post-translational modification (PTM) that plays central role in diverse physiological processes. Study on the scope and pattern of lysine acetylation is an important subject in the proteomic research. However, identification of lysine acetylation from biological sources is a great challenge due to the low abundance in massive background of unmodified proteins and the dynamic fashion of the modifications. In this research, a novel molecularly imprinted polymer (MIP) with high affinity for peptides containing acetylated lysine (Kac) has been synthesized by the combination of epitope and surface-confined imprinting strategy. A dipeptide: KacA, containing acetylated lysine and alanine residues, was used as the template and immobilized on the sacrificial silica support. After hierarchical imprinting and removal of silica, the surface-confined cavities were created on the resulting KacA-MIP material. The equilibrium binding and HPLC experiments demonstrated that the KacA-MIP has good selectivity and epitope affinity. It can differentiate Lys-acetylated peptides (Kac-peptides) from their native structures and has higher affinity for Kac-peptides with different sequences. The selectivity of the MIP was also proved by its ability in extraction of Kac-peptides from spiked histone digest and by its enrichment performance in the whole cell lysates. The study developed a method of MIP preparation with affinity for PTM peptide based on recognition of peptide side chains. It also indicated that the MIP has potential to be used as antibody mimic in the PTM analysis.

Recent advances and application of epitope imprinted materials

“…. 如图2所示, 刘震课题组 [16,17] Figure 2 (Color online) Preparation of boronate-affinity glycan-oriented surface imprinted material (a) and its recognition toward glycan, glycopeptides and glycoprotein (b) [17] 图 3 (网络版彩色)接枝表面印迹膜制备策略 [18] Figure 3 (Color online) Schematic representation of the preparation of grafted surface imprinting strategy [18] 2019 此前, 该研究组 [40,41] 还将该策略应用于乙酰化肽段的 抗原决定基印迹中. 此外, Sellergren课题组 [42,43] [45] .…”

Molecularly Imprinted Polymer Based Sensors for Medical Applications

“…MIPs have multiple advantages including easy and cost-friendly preparation, high stability, high affinity and selectivity toward template molecule. In addition, MIPs can be employed in different areas, such as sensors [13,14], enzyme mimics [15,16], solid phase extraction [17], antibody mimics [18], enantioselective [19], biomarker [20,21] protein recognition and purification [22,23], food safety [24], sensing of microorganisms [25], drug analysis [26,27,28], drug delivery system [29], hormone detoxification [30], food and environmental analysis [31]. Sensors are self-contained integrated devices which are able to convert chemical signal depending on analyte concentration into a measurable analytical signal [32,33].…”

A novel Wulff‐type boronate acid‐functionalized magnetic metal‐organic framework imprinted polymer for specific recognition of glycoproteins under physiological pH

“…Interestingly, molecular imprinting technology is an effective method to improve the specific recognition ability of materials [25]. The technique is the process of imprinting a template on the surface of a nanoparticle, and then eluting the template to leave a cavity consistent with the shape of the template molecule to obtain an imprinted polymer [26‐29]. Generally, people are committed to combining molecularly imprinted and boric acid affinity into the same unit for specific recognition of glycoproteins.…”