US 11,959,927 B2
Alpha-synuclein detection using beads
Luis Concha, San Diego, OH (US); Carly Farris, San Diego, CA (US); Bret Holguin, San Diego, CA (US); Russell Lebovitz, Oakland, CA (US); Benedikt Vollrath, San Diego, CA (US); and Frank Espin, Oceanside, CA (US)
Assigned to Amprion, Inc., San Francisco, CA (US)
Filed by Amprion, Inc., San Francisco, CA (US)
Filed on Jun. 14, 2021, as Appl. No. 17/346,854.
Application 17/346,854 is a continuation of application No. 17/011,374, filed on Sep. 3, 2020, granted, now 11,079,396.
Claims priority of provisional application 63/073,420, filed on Sep. 1, 2020.
Claims priority of provisional application 63/073,424, filed on Sep. 1, 2020.
Claims priority of provisional application 63/045,593, filed on Jun. 29, 2020.
Claims priority of provisional application 63/042,679, filed on Jun. 23, 2020.
Claims priority of provisional application 63/040,144, filed on Jun. 17, 2020.
Claims priority of provisional application 62/895,535, filed on Sep. 4, 2019.
Prior Publication US 2021/0311077 A1, Oct. 7, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. G01N 33/68 (2006.01)
CPC G01N 33/6896 (2013.01) [G01N 2800/2814 (2013.01); G01N 2800/2835 (2013.01)] 19 Claims
 
1. An in vitro method for detecting the presence of alpha-synuclein (α-syn) aggregate in a biological sample, the method comprising:
(A) providing a biological sample;
(B) providing a pre-incubation mixture, the pre-incubation mixture comprising:
(1) a monomeric α-syn protein;
(2) a buffer composition;
(3) a salt;
(4) a fluorescent protein aggregation indicator; and
(5) one or more Si3N4 beads having a diameter of 1 mm to 5 mm;
(C) combining the biological sample and the pre-incubation mixture to form an incubation mixture;
(D) incubating and agitating the incubation mixture to form an incubated mixture;
(E) illuminating the incubated mixture with a wavelength of light that excites the fluorescent protein aggregation indicator; and
(F) determining a level of fluorescence during incubation,
wherein an increase in the level of fluorescence during incubation indicates the presence of α-syn aggregate in the biological sample.