Abstract
Single-cell profiling provides insights into cellular behaviour that macroscale cell cultures and bulk measurements cannot reveal. In the context of personalized cancer treatment, the profiling of individual tumour cells may lead to higher success rates for therapies by rapidly selecting the most efficacious drugs. Currently, genomic analysis at the single-cell level is available through highly sensitive sequencing approaches. However, the identification and quantification of intracellular or secreted proteins or metabolites remains challenging. Here, we introduce a microfluidic method that facilitates capture, automated data acquisition and the multiplexed quantification of proteins from individual cells. The microfluidic platform comprises 1026 chambers with a volume of 152 pL each, in which single cells and barcoded beads are co-immobilized. We demonstrated multiplexed single-cell protein quantification with three different mammalian cell lines, including two model breast cancer cell lines. We established on-chip immunoassays for glyceraldehyde-3-phosphate dehydrogenase (GAPDH), galectin-3 (Gal-3) and galectin-3 binding protein (Gal-3bp) with detection limits as low as 7.0 × 104, 2.3 × 105 and 1.8 × 103 molecules per cell, respectively. The three investigated cell types had high cytosolic levels of GAPDH and could be clearly differentiated by their expression levels of Gal-3 and Gal-3bp, which are important factors that contribute to cancer metastasis. Because it employed commercially available barcoded beads for this study, our platform could be easily used for the single-cell protein profiling of several hundred different targets. Moreover, this versatile method is applicable to the analysis of bacteria, yeast and mammalian cells and nanometre-sized lipid vesicles.
Highlights
Cell-to-cell heterogeneity is a universal characteristic of cell populations, allowing essential processes such as cell adaptation and evolution[1]
Cellular heterogeneity plays an important role in diseases such as cancer, in which the presence of cancer cells of various subtypes can influence the progress of the disease and therapeutic success
When cellular variations in tumour tissue or between different metastatic sites can be precisely determined, the best treatment for each patient can be selected at an early stage of the disease[2]
Summary
Cell-to-cell heterogeneity is a universal characteristic of cell populations, allowing essential processes such as cell adaptation and evolution[1]. Cellular heterogeneity plays an important role in diseases such as cancer, in which the presence of cancer cells of various subtypes can influence the progress of the disease and therapeutic success. When cellular variations in tumour tissue or between different metastatic sites can be precisely determined, the best treatment for each patient can be selected at an early stage of the disease[2]. Cancer immunotherapies have been introduced that provide improved success rates and reduced side effects compared with traditional chemotherapy in certain subsets of patients[3,4]. As immunotherapies target specific biomolecules, they require. Many new immunotherapeutic drugs are currently being tested, and numerous molecules have been suggested as potential targets[7].
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