US 12,077,560 B2
Devices and methods for the sensitive detection and quantitation of biomolecules
Jennifer M. Nguyen, Uxbridge, MA (US); Matthew A. Lauber, North Smithfield, RI (US); Yun Wang Alelyunas, Stow, MA (US); Gregory T. Roman, North Scituate, RI (US); and Henry Y. Shion, Hopkinton, MA (US)
Assigned to Waters Technologies Corporation, Milford, MA (US)
Filed by Waters Technologies Corporation, Milford, MA (US)
Filed on Mar. 24, 2021, as Appl. No. 17/211,092.
Claims priority of provisional application 62/994,431, filed on Mar. 25, 2020.
Prior Publication US 2021/0300965 A1, Sep. 30, 2021
Int. Cl. B01L 3/00 (2006.01); A23C 9/146 (2006.01); B01D 53/10 (2006.01); B01F 23/00 (2022.01); B01F 23/41 (2022.01); B01F 101/23 (2022.01); B23Q 17/24 (2006.01); C07K 1/18 (2006.01); C12M 1/34 (2006.01); C12Q 1/04 (2006.01); C12Q 1/18 (2006.01); G01N 21/3577 (2014.01); G01N 21/359 (2014.01); G01N 21/39 (2006.01); G01N 21/45 (2006.01); G01N 21/64 (2006.01); G01N 30/12 (2006.01); G01N 30/68 (2006.01); G01N 30/70 (2006.01); G01N 30/72 (2006.01); G01N 30/88 (2006.01); G01N 33/00 (2006.01); G01N 33/18 (2006.01); G01N 33/50 (2006.01); G01N 33/68 (2006.01)
CPC C07K 1/18 (2013.01) [B01D 53/10 (2013.01); A23C 9/1465 (2013.01); C12Q 2547/101 (2013.01)] 13 Claims
OG exemplary drawing
 
1. A method for separating and analyzing one or more intact proteins comprising:
introducing a sample comprising the one or more intact proteins into an inlet of a chromatographic separation device that comprises
(a) sorbent particles formed from an organic polymer formed from one or more monofunctional or multifunctional aromatic monomers and having width ranging from 0.5 to 100 m and
(b) the chromatographic separation device comprising the inlet, an outlet and an internal volume between the inlet and the outlet, the internal volume being filled with a volume of the sorbent particles, wherein the internal volume has a length ranging from 2 to 50 mm and a width ranging from 50 m to 10 mm, wherein a direction of fluid flow from the inlet to the outlet is along the length of the volume of the sorbent particles, and wherein a ratio of the length to the width of the volume of the sorbent particles ranges from 10:1 to 2:1,
thereby loading the one or more intact proteins onto the sorbent particles; and introducing an acidic mobile phase comprising formic acid, acetic acid and/or a fluorinated carboxylic acid into the inlet of the chromatographic separation device in a volume sufficient to elute at least a portion of the loaded one or more intact proteins from the sorbent particles in an eluent stream from the outlet of the chromatographic separation device; and
analyzing the eluent stream with a mass spectrometer, wherein the mass spectrometer is calibrated with a lockspray standard electrosprayed under conditions which produce a lockmass ion between 2,000 and 4,000 m/z.