Metal-Chelating Peptides (MCPs), obtained from protein hydrolysates, present various applications in the field of nutrition, pharmacy, cosmetic etc. The separation of MCPs from hydrolysates mixture is challenging, yet, techniques based on peptide-metal ion interactions such as Immobilized Metal Ion Affinity Chromatography (IMAC) seem to be efficient. However, separation processes are time consuming and expensive, therefore separation prediction using chromatography modelling and simulation should be necessary. Meanwhile, the obtention of sorption isotherm for chromatography modelling is a crucial step. Thus, Surface Plasmon Resonance (SPR), a biosensor method efficient to screen MCPs in hydrolysates and with similarities to IMAC might be a good option to acquire sorption isotherm. This review highlights IMAC experimental methodology to separate MCPs and how, IMAC chromatography can be modelled using transport dispersive model and input data obtained from SPR for peptides separation simulation.

A novel approach consisting of preselection of peptides using bioinformatics tool followed by final selection using Surface Plasmon Resonance (SPR) - an efficient technique to investigate metal complexing properties of peptides/hydrolysates - was developed. Selected pea hydrolysates and synthetic metal chelating peptides potentially present in pea hydrolysates were investigated for their ability to inhibit the lipid oxidation in emulsions composed of 5 % w/w fish oil and stabilized with Tween® 20. Results indicated that addition of peptides/hydrolysates did not impact the physical stability of emulsions and led to lower level of lipid hydroperoxides. Moreover, peptide KGKSR inhibited the generation of 1-penten-3-ol and hexanal to the same level as ethylenediaminetetraacetic acid (EDTA) did and the formation of 2 ethyl-furan was lower than when EDTA was added. Peptide GRHRQKHS showed same concentration of hexanal as EDTA thus confirming efficacy of using SPR for selecting peptides/hydrolysates to use as antioxidants in emulsions.

Various peptides can be obtained through protein enzymatic hydrolysis. Immobilized metal ion affinity chromatography (IMAC) is one of the methods which can be used to separate metal chelating peptides (MCPs) in a hydrolysate mixture. In this context, this work aims to understand deeply the interactions in IMAC and surface plasmon resonance (SPR) in order to validate experimentally the analogy between both technologies and to be further able to perform IMAC modeling in the next work using peptide sorption isotherm parameters obtained from SPR. Indeed, chromatography modeling can be used to predict separation of MCPs in IMAC and the knowledge of peptide sorption isotherm obtained from SPR is a crucial step. For this purpose, 22 peptides were selected and investigated in IMAC using HisTrap X-Ni²⁺ and HiFliQ NTA-Ni²⁺ columns and were also studied in SPR as well. Results showed that peptides with histidine residues had good affinity to Ni²⁺, while the high positive charge of peptides was responsible of ionic interactions. Further, most of the peptides with good retention time in IMAC showed a good affinity in SPR as well, which validated experimentally the SPR-IMAC analogy.

Chromatography modeling for simulation is a tool that can help to predict the separation of molecules inside the column. Knowledge of sorption isotherms in chromatography modeling is a crucial step and methods such as frontal analysis or batch are used to obtain sorption isotherm parameters, but they require a significant quantity of samples. This study aims to predict Langmuir isotherm parameters from Surface Plasmon Resonance (SPR) affinity data (requiring less quantity of sample) to simulate metal chelating peptides (MCPs) separation in Immobilized Metal ion Affinity Chromatography (IMAC), thanks to the analogy between both techniques. The validity of simulation was evaluated by comparing the peptide’s simulated retention time with its experimental retention time obtained by IMAC. Results showed that the peptide affinity constant (K_A) can be conserved between SPR and IMAC. However, the maximal capacity (q_max) must be adjusted by a correction factor to overcome the geometry differences between IMAC (spherical particles) and SPR (plane sensor ship). Therefore, three approaches were studied; the best one was to use q_{max,IMAC imidazole} determined experimentally while a correction factor was applied on q_max,SPR to obtain the q_max,IMAC of the peptide, thus minimizing the discrepancy between the experimental and simulated retention times of a peptide.