US 11,808,769 B2
Spatially-tagged analyte capture agents for analyte multiplexing
Cedric Uytingco, Milpitas, CA (US); James Michael Chell, Stockholm (SE); Marlon Stoeckius, Stockholm (SE); Albert Dale Kim, Oakland, CA (US); Dulce Ovando Morales, Concord, CA (US); Andrej Hartnett, Oakland, CA (US); Yifeng Yin, Elk Grove, CA (US); and Jennifer Chew, Hayward, CA (US)
Assigned to 10x Genomics, Inc., Pleasanton, CA (US)
Filed by 10x Genomics, Inc., Pleasanton, CA (US)
Filed on Jan. 26, 2023, as Appl. No. 18/160,084.
Application 18/160,084 is a continuation of application No. 17/738,808, filed on May 6, 2022, granted, now 11,592,447.
Application 17/738,808 is a continuation of application No. PCT/US2020/049048, filed on Sep. 2, 2020.
Claims priority of provisional application 63/039,928, filed on Jun. 16, 2020.
Claims priority of provisional application 63/019,209, filed on May 1, 2020.
Claims priority of provisional application 63/000,313, filed on Mar. 26, 2020.
Claims priority of provisional application 62/959,526, filed on Jan. 10, 2020.
Claims priority of provisional application 62/939,488, filed on Nov. 22, 2019.
Claims priority of provisional application 62/933,299, filed on Nov. 8, 2019.
Prior Publication US 2023/0228762 A1, Jul. 20, 2023
Int. Cl. G01N 33/68 (2006.01); C12N 15/10 (2006.01); C12Q 1/6804 (2018.01); C12Q 1/6874 (2018.01); G01N 1/30 (2006.01); G01N 21/64 (2006.01); G01N 33/58 (2006.01)
CPC G01N 33/6803 (2013.01) [C12N 15/1065 (2013.01); C12Q 1/6804 (2013.01); C12Q 1/6874 (2013.01); G01N 1/30 (2013.01); G01N 21/6428 (2013.01); G01N 21/6456 (2013.01); G01N 33/582 (2013.01); G01N 2021/6439 (2013.01)] 26 Claims
 
1. A method for preparing a biological sample for spatial proteomic analysis, the method comprising:
(a) providing a substrate comprising a plurality of capture probes, wherein a capture probe of the plurality of capture probes comprises a spatial barcode and a capture domain;
(b) providing the biological sample on a second substrate;
(c) aligning the biological sample on the second substrate with the substrate comprising the plurality of capture probes, such that at least a portion of the biological sample is aligned with at least a portion of the substrate comprising the plurality of capture probes;
(d) contacting the biological sample with a protein blocking buffer; and
(e) contacting the biological sample with a plurality of analyte capture agents, wherein an analyte capture agent of the plurality of analyte capture agents comprises:
(i) an analyte binding moiety that binds to a protein from the biological sample;
(ii) an analyte binding moiety barcode; and
(iii) an analyte capture sequence, wherein the analyte capture sequence hybridizes to the capture domain of the capture probe.