Proteases and Cancer Methods and Protocols

Web-based resources to investigate protease function.- N-terminomics/TAILS of human cancer biopsies and cancer cell lines.- A new affinity-based probe to profile MMP active forms.- N-glycan insertion for probing protein-protein interactions and epitope mapping.- Expression and purification of recombinant ADAMTS8.- Expression and purification of active monomeric MMP7.- Mapping the binding sites of MMPs on type II and III collagens using triple-helical peptide Toolkits.- Determination of Versikine Levels by Enzyme-Linked Immunosorbent Assay (ELISA).- Methods to investigate thrombin cleavage of osteopontin (OPN).- Using siRNA silencing to analyze ADAM17 in macrophages.- Preparation and characterization of collagen-hyaluronic acid (Col-HA) matrices: in vitro mimics of the tumor microenvironment.- Assessment of Invadopodium Formation and Gelatin Degradation In Vitro.- Investigation of MT1-MMP activity in cancer cells.- Gelatinin situ zymography to study gelatinase activity in colon cancer cells treated with platelet microparticles (PMPs).- Interrogating the impact of protease activity on tumor progression using 3D spheroid models.- Analysis of Matrix Metalloproteinase Activity and Inhibition in Cancer Spheroids.- Intravital microscopy to study the effect of matrix metalloproteinases inhibition on Acute myeloid leukemia cells migration in the bone marrow.- Fluorescence-based peptidolytic assay for high throughput screening of MMP14 inhibitors.- Generation of protease inhibitory antibodies by functional in vivo selection.- Engineering selective TIMPs using a counter-selective screening strategy.- Engineering new protease inhibitors using a2-macroglobulin.- Design of bioengineered peptides/proteases as anti-cancer reagents with integrated omics and machine-learning approaches.