Chemical Crosslinking‐Mass Spectrometry (CXL‐MS) for Proteomics, Antibody‐Drug Conjugates (ADCs) and Cryo‐Electron Microscopy (cryo‐EM)

Abstract

Chemical crosslinking-mass spectrometry (CXL-MS) has emerged as a powerful tool in structural biology to elucidate protein-protein interactions. This review throws light on the advent of these technologies incorporating chemical crosslinking in proteomics, structural biology and extended to studies and applications of antibody-drug conjugates (ADCs). Monoclonal ASCs are known to target their specific receptors, where the drug could be released for vastly improved therapeutic effect. Cryo-electron microscopy (cryo-EM) is being referred to as the "revolution of the century" as it can help attain almost atomic resolution, while preserving the biological samples at near-native state and has been recognized by the Nobel award 2017. CXL-MS and appropriate bioinformatics tools in combination with cryo-EM reveal better 3D information about dynamic interactions in proteins, interactive domains and motifs, among others in ADCs and in viruses like Zika virus, Rota virus, Dengue virus and also spliceosomes at resolutions, especially below 3 Å. © 2018 IUBMB Life, 70(10):947-960, 2018.

A Simple Cross-Linking/Mass Spectrometry Workflow for Studying System-wide Protein Interactions

“…The resulting map of amino acid distances can be computationally reprocessed resulting in 3D structural models of the proteins or protein assemblies. − The main advantage of XLMS over other protein structural techniques is that it creates comprehensive snapshots of the protein landscape with minimum interference. XLMS experiments can be conducted within a few days, making XLMS a highly attractive approach that complements existing high-resolution protein structural techniques, such as nuclear magnetic resonance spectroscopy, X-ray crystallography, and cryo-electron microscopy (cryo-EM). − On top of that, XLMS requires only minute amounts of protein and can even be applied to intact cells. , System-wide XLMS offers two key benefits. (i) It allows system-wide protein interactions to be captured for a comprehensive understanding of, e.g., cellular signaling pathways, and (ii) it allows analysis of the conformation and the interaction of proteins in their native environment.…”

Antibody-Drug Conjugates, the Combination of Large and Small Therapeutic Molecules

“…17 . Z dalších potenciálních metod přichází v úvahu NMR spektroskopie v roztoku 18 a kryoelektronová mikroskopie 19 , ale tyto metody jsou většinou použity v tandemu s MS, protože jejich rozlišení je obecně nižší 20 .…”

Antibody‐Incorporated Nanomedicines for Cancer Therapy

“…[ 208 ] Crosslinkers composed of two same or different functional groups are referred to as homo‐ or heterobifunctional crosslinking reagents, respectively. [ 209 ] For example, homobifunctional crosslinker glutaraldehyde (GLU) allows firm conjugation between the aldehyde groups of GLU and the amino groups of antibodies or nanoparticles. [ 210–212 ] Aggarwal et al functionalized Gem‐loaded PLGA‐PEG‐NH 2 nanoparticles with anti‐EGFR mAbs through the GLU‐mediated crosslinking method, and the resultant product allowed targeted delivery of chemotherapeutic agents for pancreatic cancer treatment.…”