Biomimetic Affinity Ligands for Protein Purification

Abstract

The development of sophisticated molecular modeling software and new bioinformatic tools, as well as the emergence of data banks containing detailed information about a huge number of proteins, enabled the de novo intelligent design of synthetic affinity ligands. Such synthetic compounds can be tailored to mimic natural biological recognition motifs or to interact with key surface-exposed residues on target proteins and are designated as "biomimetic ligands." A well-established methodology for generating biomimetic or synthetic affinity ligands integrates rational design with combinatorial solid-phase synthesis and screening, using the triazine scaffold and analogues of amino acids side chains to create molecular diversity.Triazine-based synthetic ligands are nontoxic, low-cost, highly stable compounds that can replace advantageously natural biological ligands in the purification of proteins by affinity-based methodologies.

Materials Functionalization with Multicomponent Reactions: State of the Art

“…Of late, there has been resurgence of interest in the construction of fully synthetic low-molecular weight affinity ligands from small-molecule screening methodologies instead of biological ligands that suffered from instability, high production costs and low reusability . In a very interesting work in 2009, Lowe et al profited from the outstanding features of MCRs; being “one-pot” and high-structural variations due their inherent combinatorial nature, to combining with chromatographic solid supports for rational design of spectrum of beneficial affinity ligands-modified sepharose supports via Ugi-4CR that are suitable for the purification of immunoglobulins and their fragments by affinity chromatography .…”

One-Step Purification of Recombinant Cutinase from an E. coli Extract Using a Stabilizing Triazine-Scaffolded Synthetic Affinity Ligand

“…2.4. Solid-Phase Synthesis of Ligand 11/3 ′ Ligand 11/3 ′ was synthesized by a well-established solid-phase synthesis protocol, as described in [28]. Sepharose CL-6B (support matrix) was activated with epichlorohydrin, followed by amination and reaction with cyanuric chloride.…”

“…However, its use instead of classic chromatographic techniques is often limited by the availability of a cost-effective ligand recognizing the protein of interest. Affinity ligands for protein purification (as well as for other purposes that take advantage of affinity interactions) can be natural molecules, such as enzyme substrates and inhibitors, effectors, coenzymes, hormones, antigens, nucleic acids and sugars, for example, [21] or bioinspired affinity ligands [22] such as short mimetic peptides [23], aptamers [24], or rationally designed biomimetic triazine-scaffolded affinity ligands [25][26][27][28]. Triazine-based synthetic ligands are nontoxic, highly stable, cost-effective, low-molecular weight compounds that can replace advantageously natural biological ligands in the purification of proteins by affinity chromatographic methods [27,28].…”

“…Affinity ligands for protein purification (as well as for other purposes that take advantage of affinity interactions) can be natural molecules, such as enzyme substrates and inhibitors, effectors, coenzymes, hormones, antigens, nucleic acids and sugars, for example, [21] or bioinspired affinity ligands [22] such as short mimetic peptides [23], aptamers [24], or rationally designed biomimetic triazine-scaffolded affinity ligands [25][26][27][28]. Triazine-based synthetic ligands are nontoxic, highly stable, cost-effective, low-molecular weight compounds that can replace advantageously natural biological ligands in the purification of proteins by affinity chromatographic methods [27,28]. These synthetic ligands mimic the structure and binding of natural ligands and are easily synthetized by a solid-phase assembly method on agarose matrix with a triazine scaffold.…”

“…These synthetic ligands mimic the structure and binding of natural ligands and are easily synthetized by a solid-phase assembly method on agarose matrix with a triazine scaffold. The triazine scaffold allows for the display of attached functional groups and the possibility of attaining molecular diversity by serial substitution of the chlorines in the triazine ring by commercially available aliphatic and aromatic amines [28].…”