title: Antibody-based affinity cryo-EM grid.
abstract: The Affinity Grid technique combines sample purification and cryo-Electron Microscopy (cryo-EM) grid preparation into a single step. Several types of affinity surfaces, including functionalized lipids monolayers, streptavidin 2D crystals, and covalently functionalized carbon surfaces have been reported. More recently, we presented a new affinity cryo-EM approach, cryo-SPIEM, which applies the traditional Solid Phase Immune Electron Microscopy (SPIEM) technique to cryo-EM. This approach significantly simplifies the preparation of affinity grids and directly works with native macromolecular complexes without need of target modifications. With wide availability of high affinity and high specificity antibodies, the antibody-based affinity grid would enable cryo-EM studies of the native samples directly from cell cultures, targets of low abundance, and unstable or short-lived intermediate states.
authors.affiliation: Markey Center for Structural Biology, Department of Biological Sciences, Purdue University, West Lafayette, IN, USA.
authors.forename: Guimei
authors.initials: G
authors.lastname: Yu
authors.affiliation: Markey Center for Structural Biology, Department of Biological Sciences, Purdue University, West Lafayette, IN, USA.
authors.forename: Kunpeng
authors.initials: K
authors.lastname: Li
authors.affiliation: Markey Center for Structural Biology, Department of Biological Sciences, Purdue University, West Lafayette, IN, USA. Electronic address: jiang12@purdue.edu.
authors.forename: Wen
authors.initials: W
authors.lastname: Jiang
mesh_terms: D055910:Antibodies, Immobilized / Q000737:chemistry; D020285:Cryoelectron Microscopy / Q000379:methods*; D006801:Humans; D021621:Imaging, Three-Dimensional; D046911:Macromolecular Substances / Q000737:chemistry / Q000302:isolation & purification / Q000648:ultrastructure; D046529:Microscopy, Electron, Transmission / Q000379:methods; D011485:Protein Binding; D014771:Virion / Q000737:chemistry / Q000302:isolation & purification / Q000648:ultrastructure
chemical_list: D055910:Antibodies, Immobilized; D046911:Macromolecular Substances
keywords: 3D reconstruction; Affinity grid; Antibodies; Cryo-SPIEM; On-grid assays; Single particle cryo-EM