Full Description
This volume details bioinformatics analysis, biochemical assays, recombinant protein expression and purification, methods to investigate protease activity in cell-based, organoids and in vivo systems, proteomics, transcriptomics, machine learning and novel approaches to target dysregulated protease activity in cancer. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Cancer Immunotherapy: Methods and Protocols aims to provide the scientific community with a diverse and representative collection of state-of-the-art methods to investigate the protean functions of proteases in cancer biology.
Contents
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 α2-macroglobulin.- Design of bioengineered peptides/proteases as anti-cancer reagents with integrated omics and machine-learning approaches.
