US 12,265,089 B2
Discriminating Parkinson's disease from multiple system atrophy using alpha-synuclein PMCA
Claudio Soto Jara, Friendswood, TX (US); Mohammad Shahnawaz, Houston, TX (US); and Luis Concha, San Diego, CA (US)
Assigned to Board of Regents of the University of Texas System, Austin, TX (US); and Amprion, Inc., San Francisco, CA (US)
Filed by Board of Regents of the University of Texas System, Austin, TX (US); and Amprion, Inc., San Francisco, CA (US)
Filed on Jan. 21, 2021, as Appl. No. 17/154,966.
Application 17/154,966 is a continuation in part of application No. 17/011,374, filed on Sep. 3, 2020, granted, now 11,079,396.
Application 17/154,966 is a continuation in part of application No. 14/852,475, filed on Sep. 11, 2015, granted, now 10,989,718.
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 62/963,805, filed on Jan. 21, 2020.
Claims priority of provisional application 62/895,535, filed on Sep. 4, 2019.
Claims priority of provisional application 62/049,304, filed on Sep. 11, 2014.
Prior Publication US 2021/0164998 A1, Jun. 3, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. G01N 33/68 (2006.01); A61B 8/13 (2006.01); G01N 33/58 (2006.01)
CPC G01N 33/6893 (2013.01) [A61B 8/13 (2013.01); G01N 33/582 (2013.01); G01N 2800/2835 (2013.01); G01N 2800/2878 (2013.01)] 43 Claims
 
1. A method for differentiating a diagnosis of Parkinson's Disease (PD) from a diagnosis of Multiple System Atrophy (MSA) in a human subject, the method comprising:
(A) contacting a cerebrospinal fluid (CSF) or a brain homogenate (BH) sample (i) obtained from the human subject and (ii) comprising soluble, misfolded alpha-synuclein (αS) protein, with a pre-incubation mixture, the pre-incubation mixture comprising:
(1) a monomeric αS substrate;
(2) a buffer composition;
(3) sodium chloride (NaCl); and
(4) a fluorescent protein aggregation indicator,
to form an incubation mixture;
(B) incubating and agitating the incubation mixture to form an incubated mixture comprising misfolded αS aggregates;
(C) illuminating the incubated mixture with a wavelength of light that excites the fluorescent protein aggregation indicator, including:
(1) determining an initial average fluorescence (AFi) of the incubated mixture prior to the formation of the misfolded αS aggregates;
(2) determining a standard deviation (SDi) of the AFi; and
(3) determining a maximum fluorescence (Fmax) of the incubated mixture after the formation of the misfolded αS aggregates; and
(D) diagnosing the human subject as having PD or MSA, where:
(1) if the Fmax is from about 22.2×SDi+AFi to 250×SDi+AFi, the patient is diagnosed as having MSA; and
(2) if the Fmax is above 250×SDi+AFi, the patient is diagnosed as having PD.