US 12,203,939 B2
Quantitative massively parallel proteomics
Ian Peikon, Bethpage, NY (US); Gary Tong, Berkeley, CA (US); Daniel Liu, Berkeley, CA (US); and Ci Chu, Palo Alto, CA (US)
Assigned to Verily Life Sciences LLC, Dallas, TX (US)
Filed by Verily Life Sciences LLC, South San Francisco, CA (US)
Filed on Oct. 19, 2020, as Appl. No. 17/074,224.
Application 17/074,224 is a continuation of application No. 15/656,423, filed on Jul. 21, 2017, granted, now 10,809,266.
Claims priority of provisional application 62/365,542, filed on Jul. 22, 2016.
Prior Publication US 2021/0132078 A1, May 6, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. G01N 33/68 (2006.01); C12Q 1/6804 (2018.01); G01N 33/58 (2006.01)
CPC G01N 33/6845 (2013.01) [C12Q 1/6804 (2013.01); G01N 33/58 (2013.01); G01N 2458/10 (2013.01); G01N 2570/00 (2013.01)] 9 Claims
 
1. A method of identifying an expression profile in a single cell of a population of cells, comprising:
a) contacting a population of cells with a plurality of antibodies under conditions that promote specific binding of the antibodies to target antigens of the cells, wherein each antibody in the plurality is capable of binding to a unique target antigen in one or more cells, wherein each antibody is conjugated to an oligonucleotide via thiol/maleimide conjugation, and wherein the oligonucleotide comprises a unique molecular identifier sequence and an antibody identifier sequence corresponding to the unique target antigen;
b) separating single cells of the population into individual compartments, wherein each compartment further comprises an oligonucleotide comprising a unique cellular identifier sequence that corresponds to the single cell in each compartment;
c) lysing the single cells with an agent that extracts proteins and nucleic acids from the cells, wherein the nucleic acids comprise RNA and/or DNA;
d) amplifying the unique molecular identifier sequences and antibody identifier sequences in each individual compartment, using the oligonucleotide comprising the unique cellular identifier sequence;
e) analyzing the amplified unique molecular identifier sequences and antibody identifier sequences to identify the expression profile of each single cell, wherein the unique molecular identifier sequences reduce bias in the analysis; and
f) analyzing the RNA extracted from the cells to identify a transcriptomic profile of each single cell and/or the DNA extracted from the cells to obtain genomic information of each single cell.