Highly multiplexed and quantitative cell-surface protein profiling using genetically barcoded antibodies.

Abstract:

:Human cells express thousands of different surface proteins that can be used for cell classification, or to distinguish healthy and disease conditions. A method capable of profiling a substantial fraction of the surface proteome simultaneously and inexpensively would enable more accurate and complete classification of cell states. We present a highly multiplexed and quantitative surface proteomic method using genetically barcoded antibodies called phage-antibody next-generation sequencing (PhaNGS). Using 144 preselected antibodies displayed on filamentous phage (Fab-phage) against 44 receptor targets, we assess changes in B cell surface proteins after the development of drug resistance in a patient with acute lymphoblastic leukemia (ALL) and in adaptation to oncogene expression in a Myc-inducible Burkitt lymphoma model. We further show PhaNGS can be applied at the single-cell level. Our results reveal that a common set of proteins including FLT3, NCR3LG1, and ROR1 dominate the response to similar oncogenic perturbations in B cells. Linking high-affinity, selective, genetically encoded binders to NGS enables direct and highly multiplexed protein detection, comparable to RNA-sequencing for mRNA. PhaNGS has the potential to profile a substantial fraction of the surface proteome simultaneously and inexpensively to enable more accurate and complete classification of cell states.

authors

Pollock SB,Hu A,Mou Y,Martinko AJ,Julien O,Hornsby M,Ploder L,Adams JJ,Geng H,Müschen M,Sidhu SS,Moffat J,Wells JA

doi

10.1073/pnas.1721899115

subject

Has Abstract

pub_date

2018-03-13 00:00:00

pages

2836-2841

issue

11

eissn

0027-8424

issn

1091-6490

pii

1721899115

journal_volume

115

pub_type

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