	US 12,332,205 B2
	Droplet-based surface modification and washing
Vamsee K. Pamula, Cary, NC (US); Vijay Srinivasan, Cary, NC (US); Michael G. Pollack, Durham, NC (US); and Richard B. Fair, Durham, NC (US)
Assigned to Advanced Liquid Logic, Inc., San Diego, CA (US); and Duke University, Durham, NC (US)
Filed by Advanced Liquid Logic, Inc., San Diego, CA (US); and Duke University, Durham, NC (US)
Filed on Feb. 3, 2022, as Appl. No. 17/649,842.
Application 12/113,385 is a division of application No. 11/639,736, filed on Dec. 15, 2006, granted, now 7,439,014, issued on Oct. 21, 2008.
Application 17/649,842 is a continuation of application No. 16/788,187, filed on Feb. 11, 2020, granted, now 11,255,809.
Application 16/788,187 is a continuation of application No. 15/099,147, filed on Apr. 14, 2016, abandoned.
Application 15/099,147 is a continuation of application No. 14/609,744, filed on Jan. 30, 2015, abandoned.
Application 14/609,744 is a continuation of application No. 12/615,666, filed on Nov. 10, 2009, granted, now 8,951,721, issued on Feb. 10, 2015.
Application 12/615,666 is a continuation of application No. 12/113,385, filed on May 1, 2008, granted, now 8,541,176, issued on Sep. 24, 2013.
Claims priority of provisional application 60/807,104, filed on Jul. 12, 2006.
Claims priority of provisional application 60/806,412, filed on Jun. 30, 2006.
Claims priority of provisional application 60/746,797, filed on May 9, 2006.
Claims priority of provisional application 60/746,801, filed on May 9, 2006.
Claims priority of provisional application 60/745,914, filed on Apr. 28, 2006.
Claims priority of provisional application 60/745,950, filed on Apr. 28, 2006.
Claims priority of provisional application 60/745,059, filed on Apr. 18, 2006.
Claims priority of provisional application 60/745,058, filed on Apr. 18, 2006.
Claims priority of provisional application 60/745,043, filed on Apr. 18, 2006.
Prior Publication US 2022/0155251 A1, May 19, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. G01N 27/26 (2006.01); B01F 33/302 (2022.01); B01F 33/3031 (2022.01); B01L 3/00 (2006.01); B01L 7/00 (2006.01); C12Q 1/686 (2018.01); C12Q 1/6869 (2018.01); G01N 27/327 (2006.01); G01N 27/416 (2006.01); G01N 27/447 (2006.01); G01N 35/00 (2006.01); B01L 3/02 (2006.01)

CPC G01N 27/26 (2013.01) [B01F 33/3021 (2022.01); B01F 33/3031 (2022.01); B01L 3/502715 (2013.01); B01L 3/502723 (2013.01); B01L 3/502761 (2013.01); B01L 3/502784 (2013.01); B01L 3/502792 (2013.01); B01L 7/525 (2013.01); C12Q 1/686 (2013.01); C12Q 1/6869 (2013.01); G01N 27/3271 (2013.01); G01N 27/416 (2013.01); G01N 27/447 (2013.01); G01N 35/00029 (2013.01); B01L 3/0268 (2013.01); B01L 7/52 (2013.01); B01L 2200/027 (2013.01); B01L 2200/0668 (2013.01); B01L 2200/0673 (2013.01); B01L 2200/0684 (2013.01); B01L 2300/0654 (2013.01); B01L 2300/0816 (2013.01); B01L 2300/0861 (2013.01); B01L 2300/0864 (2013.01); B01L 2300/0867 (2013.01); B01L 2300/089 (2013.01); B01L 2300/1827 (2013.01); B01L 2400/0406 (2013.01); B01L 2400/0415 (2013.01); B01L 2400/0424 (2013.01); B01L 2400/0427 (2013.01); B01L 2400/043 (2013.01); B01L 2400/0436 (2013.01); B01L 2400/0442 (2013.01); B01L 2400/0448 (2013.01); G01N 2035/00158 (2013.01); G01N 2035/00237 (2013.01)]

15 Claims

1. A method comprising:

immobilizing on a surface an antibody with specificity for a target analyte, wherein the surface is a surface of one or more beads;

washing the immobilized antibody to remove excess antibody;

using electrowetting droplet operations of two parallel substrates separated by a gap to expose the immobilized antibody to a sample droplet, where one or both of the substrates comprise electrodes, where the electrodes are capable of conducting one or more droplet operations, where the electrodes are electronically coupled to and controlled by a controller;

using electrowetting droplet operations to wash the immobilized antibody to remove unbound components of the sample droplet;

using electrowetting droplet operations to expose the immobilized antibody to a droplet including a reporter antibody;

using electrowetting droplet operations to wash away excess reporter antibody; and

measuring a measurable parameter or signal facilitated by the reporter antibody.