Trypsin immobilization on three monolithic disks for on-line protein digestion

Total on-line analysis of a target protein from plasma by immunoextraction, digestion and liquid chromatography–mass spectrometry

“…Trypsin based IMERs were already successfully developed under various formats, i.e. pre-columns [17][18][19][20][21][22][23], membranes [24], capillaries [25][26][27][28][29] and disks [30][31][32]. Recently, our group highlighted the important effect of the immobilization support on the performances of the resulting IMER [33].…”

On-line chromatographic screening of matrix metalloproteinase inhibitors using immobilized MMP-9 enzyme reactor

“…Identification of potential leads from combinatorial libraries demands a high throughput screening (HTS) platform that can select potential effectors of pharmacological activity against disease targets for further optimization and development. While the multiwell robotic screening platform has revolutionized HTS in many areas, other ligand-based HTS methodologies, such as the "target protein-immobilized" flow-through systems are less explored [1][2][3]. Enzymes are versatile catalysts that mediate diversified metabolic and pharmacological processes in a biological system.…”

“…Immobilization of enzymes is therefore explored as an attempt to stabilize and prolong the activity and reusability of the catalyst [5]. In the last 3 years, immobilized enzyme reactors (IMERs) coupled to separation modules have been developed to screen enzyme inhibitors [6,7], profile drug metabolism [8], map proteins [1,9], and analyse enantiomers [10] in a rapid and automatic fashion.…”

The Value of Fungal Protease Inhibitors in Affinity Chromatography

“…This method is faster than the epoxy method and involves fewer steps (Mallik & Hage, 2006). In addition, the remaining free imidazole groups of the supports are rapidly selfdeactivated after the immobilization process in aqueous solution, forming the original hydroxyl groups and releasing CO 2 and imidazole (Benčina et al, 2004;Nicoli et al, 2008).…”