US 12,287,331 B2
Compositions, methods and systems for protein corona analysis and uses thereof
Hongwei Xia, Fremont, CA (US); Lyndal Hesterberg, Loveland, CO (US); Michael Figa, San Mateo, CA (US); Xiaoyan Zhao, Foster City, CA (US); Gregory Troiano, Pembroke, MA (US); William Manning, Redwood City, CA (US); John Blume, Bellingham, WA (US); Omid Farokhzad, Waban, MA (US); Matthew McLean, Redwood City, CA (US); Craig Stolarczyk, San Mateo, CA (US); Marwin Ko, Redwood City, CA (US); and Theodore Platt, Danville, CA (US)
Assigned to Seer, Inc., Redwood City, CA (US)
Filed by Seer, Inc., Redwood City, CA (US)
Filed on Sep. 1, 2023, as Appl. No. 18/460,474.
Application 18/460,474 is a continuation of application No. 17/216,514, filed on Mar. 29, 2021.
Application 17/216,514 is a continuation of application No. PCT/US2019/000061, filed on Nov. 7, 2019.
Claims priority of provisional application 62/874,862, filed on Jul. 16, 2019.
Claims priority of provisional application 62/824,278, filed on Mar. 26, 2019.
Claims priority of provisional application 62/756,960, filed on Nov. 7, 2018.
Prior Publication US 2023/0417744 A1, Dec. 28, 2023
This patent is subject to a terminal disclaimer.
Int. Cl. G01N 33/543 (2006.01); G01N 33/551 (2006.01); G01N 33/68 (2006.01)
CPC G01N 33/54326 (2013.01) [G01N 33/54346 (2013.01); G01N 33/551 (2013.01); G01N 33/6848 (2013.01)] 28 Claims
OG exemplary drawing
 
1. A high-throughput method of preparing to identify proteins in a biological sample, the method comprising:
(a) incubating magnetic microparticles with the biological sample to form protein coronas comprising proteins, the magnetic microparticles comprising: (i) silica, (ii) carboxylate functional groups, (iii) negative surface charges from −30 mV to −10 mV in zeta potential, wherein the zeta potential is measurable with a concentration of 0.02 wt % in a solution in 5% pH 7.4 PBS at about 25° C. with an about 1 minute equilibration time on a Zetasizer Nano ZS (Malvern Instruments) in the absence of biological sample, and (iv) a heterogeneous distribution of sizes including from about 1 μm to about 10 μm in diameter, wherein the magnetic microparticles are configured to form protein coronas having at least 200 unique protein groups that can be identified using mass spectrometry;
(b) magnetically isolating the magnetic microparticles, by applying an external magnetic field, from unbound protein in the biological sample to enrich the proteins in the protein coronas;
(c) washing the enriched proteins and the magnetic microparticles after magnetically isolating the magnetic microparticles;
(d) incubating the enriched proteins in the protein coronas by adding a denaturing reagent to the protein coronas and the magnetic microparticles;
(e) digesting the enriched proteins in the protein coronas to generate digested peptides by adding a digestion reagent to the protein coronas and the magnetic microparticles; and
(f) purifying the digested peptides.