Antibody-Based Affinity Cryoelectron Microscopy at 2.6-Å Resolution.
The affinity cryoelectron microscopy (cryo-EM) approach has been explored in recent years to simplify and/or improve the sample preparation for cryo-EM, which can bring previously challenging specimens such as those of low abundance and/or unpurified ones within reach of the cryo-EM technique. Despite the demonstrated successes for solving structures to low to intermediate resolutions, the lack of near-atomic structures using this approach has led to a common perception of affinity cryo-EM as a niche technique incapable of reaching high resolutions. Here, we report a ∼2.6-Å structure solved using the antibody-based affinity grid approach with low-concentration Tulane virus purified from a low-yield cell-culture system that has been challenging to standard cryo-EM grid preparation. Quantitative analyses of the structure indicate data and reconstruction quality comparable with the conventional grid preparation method using samples at high concentration.
Department of Biological Science, Markey Center for Structural Biology, Purdue University, West Lafayette, IN 47907, USA.
Guimei
G
Yu
Department of Biological Science, Markey Center for Structural Biology, Purdue University, West Lafayette, IN 47907, USA.
Kunpeng
K
Li
Divisions of Infectious Diseases, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA; Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, OH 45267, USA.
Pengwei
P
Huang
Divisions of Infectious Diseases, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA; Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, OH 45267, USA.
Xi
X
Jiang
Department of Biological Science, Markey Center for Structural Biology, Purdue University, West Lafayette, IN 47907, USA. Electronic address: jiang12@purdue.edu.
Wen
W
Jiang
D000906:Antibodies / Q000378:metabolism*; D020285:Cryoelectron Microscopy / Q000379:methods*; D008958:Models, Molecular; D015394:Molecular Structure; D014780:Viruses / Q000737:chemistry* / Q000276:immunology
D000906:Antibodies
Tulane virus; affinity cryoelectron microscopy; antibody-based affinity grid; single-particle 3D reconstruction