US 12,347,524 B2
Quantum mechanical X-ray crystallography and Cryo-EM diagnostic for molecules
Lance Michael Westerhoff, Annville, PA (US); Oleh Y. Borbulevych, Bellefonte, PA (US); and Roger Isaac Martin, State College, PA (US)
Assigned to QuantumBio Inc., State College, PA (US)
Filed by QuantumBio Inc., State College, PA (US)
Filed on Jun. 15, 2021, as Appl. No. 17/348,074.
Application 16/801,588 is a division of application No. 15/017,453, filed on Feb. 5, 2016, granted, now 10,614,909, issued on Apr. 7, 2020.
Application 17/348,074 is a continuation in part of application No. 16/801,588, filed on Feb. 26, 2020, granted, now 11,688,488.
Claims priority of provisional application 62/157,787, filed on May 6, 2015.
Claims priority of provisional application 62/112,951, filed on Feb. 6, 2015.
Prior Publication US 2021/0407620 A1, Dec. 30, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. G01N 33/48 (2006.01); G01N 23/20008 (2018.01); G01N 33/50 (2006.01); G01N 33/68 (2006.01); G16B 15/00 (2019.01); G16B 20/00 (2019.01); G16B 20/30 (2019.01)
CPC G16B 15/00 (2019.02) [G01N 23/20008 (2013.01); G01N 33/6803 (2013.01); G16B 20/00 (2019.02); G16B 20/30 (2019.02); G01N 2223/304 (2013.01); G01N 2223/612 (2013.01)] 8 Claims
 
1. An analytic method more efficiently to identify conformational, protonation, or solvent effect information from a molecule of interest by using cryogenic electron microscopy (Cryo-EM), comprising the steps of a) selecting an aliquot of a molecule sample as a sample to be diagnosed; b) imaging said molecule by Cryo-EM and collecting a quantity of density data generated thereby; c) assaying Cryo-EM density within said density data and creating from said density data thus assayed a population set containing a plurality of set elements consisting of all or reasonably all possible protomer/tautomer and conformational states of said molecule; d) determining a local ligand strain energy value SE for each of said elements; e) determining ZDD for each of said elements by calculating a difference density Z for each element and compiling ZDD data therefrom: and f) selecting a single element from among said elements that represents the true state of at least one moiety of said molecule by calculating Scorei according to the following equation,
Scorei={((ZDDi−uZDD)/σZDD)+((SEi−μSE)/σSE)}
wherein the highest Scorei obtained for said population corresponds to the best form “i” that fits both SE and ZDD criteria, so that Scorei when output to a user identifies the element from said population that most closely corresponds with said molecule thus diagnosed with increased time efficiency using said low resolution Cryo-EM compared to the same method using atomic resolution x-ray crystallography or Cryo-EM.