Biomarker Detection Methods and Systems and Kits for Practicing Same

Abstract

Aspects of the present disclosure include methods that include co-culturing a cell and a microparticle that includes a capture ligand, in a culture medium under conditions in which a biomarker produced by the cell is bound by the capture ligand. Such methods may further include detecting (e.g., by flow or mass cytometry) complexes that include the microparticle, the capture ligand, the biomarker, and a detection reagent. The methods may further include determining the proportion or number of cells among a heterogeneous cell population that produced the biomarker and/or the level of biomarker secreted by such cells. Compositions, systems and kits are also provided.

1 .- 55 . (canceled) 
     
     
         56 . A system, comprising:
 a processor; and   a non-transitory computer readable medium comprising instructions that cause the processor to:   count a number of positive microparticle complexes, wherein the positive microparticle complexes comprise a microparticle, a capture ligand, a biomarker, and a fluorescently-labeled detection reagent;   determine the total number of microparticle complexes acquired by the system;   calculate the percentage of positive microparticle complexes among the total number of microparticle complexes; and   determine the number and/or proportion of cells in a cell-microparticle co-culture that comprised the biomarker.   
     
     
         57 . The system of  claim 56 , wherein the non-transitory computer readable medium further comprises instructions that cause the processor to:
 determine a mean or a median fluorescence intensity of the positive microparticle complexes acquired by the system; and   determine a level of the biomarker present in the cell-microparticle co-culture.   
     
     
         58 . The system of  claim 56 , wherein the system is a flow cytometry system or a Luminex detecting system. 
     
     
         59 . The system of  claim 56 , wherein the system is a mass cytometry system. 
     
     
         60 .- 68 . (canceled)