Simultaneous Detection of cfDNA and Exosomal Biomarkers

Expanding biomarker research capabilities

Accelerate and enhance your biomarker discovery and characterization studies with the XCF™ COMPLETE Exosome & cfDNA Isolation Kit. This unique, two-in-one product delivers simultaneous isolation of cell free DNA (cfDNA) and exosomal DNA from the same sample, streamlining sample processing and reducing the workload of separately isolating DNA from multiple sources. In addition, the isolated exosomes can also be analyzed for protein, lipid, miRNA, and metabolite content, efficiently broadening your biomarker discovery capabilities and enabling correlation and co-analysis of cfDNA biomarkers with exosomal biomarkers.

With the growing recognition that circulating, cfDNA can provide critical information on diseases that are normally difficult to detect with standard approaches—including cancer and neurological disorders—the interest in liquid biopsy methods for biomarker discovery and disease research is on the rise. However, in addition to cfDNA, exosomes are also a rich source of biomarkers that are amenable to the straightforward liquid biopsy approach. To help scientists maximize their biomarker discovery, SBI has developed the XCF COMPLETE Exosome & cfDNA Isolation Kit, which enables quick and easy isolation of both exosomes or exosomal DNA (in one aliquot) and cfDNA (in a separate aliquot) from a single serum or plasma sample.

• Comprehensive—with a single kit you can obtain two distinct sources of biomarkers from a single serum or plasma sample
• Time-saving—requires less than thirty minutes of hands-on time
• Fully compatible—isolated DNA is compatible with downstream applications, such as qPCR and NGS
• Clean—minimal carryover of co-precipitating proteins, such as albumin and IgG, for good performance in sensitive downstream applications
• Flexible—choose the XCF COMPLETE Exosome & cfDNA Isolation Kit which includes ExoQuick for exosome and exosomal DNA isolation, or the XCF Exosomal DNA Isolation Kit if you already have purified exosomes

Figure 1. Accelerate your biomarker research with simultaneous isolation of exosomal DNA and cfDNA. Note that other types of exosome-derived proteins, miRNA, lipids, and more can be analyzed as well.

Validation

Demonstrably high performance for biomarker research

SBI’s XCF™ COMPLETE Exosome & cfDNA Isolation Kit delivers high performance in a quick and easy workflow. Samples processed with the reagents in this kit show much lower protein carryover than untreated samples (Figure 1), the expected exosome particle size profile (Figure 2), excellent exosomal and cfDNA yields and quality (Figure 3), and the expected DNA sizes for both exosomal DNA and cfDNA (Figure 3).

Figure 1. The XCF COMPLETE Kit shows low protein carryover. Western blots of co-purifying human albumin (~67kD) and heavy-chain IgG (~50kD) comparing untreated serum and plasma (left two lanes) versus the same samples processed using the XCF Kit (right two lanes), show much less protein carryover in the samples processed using the XCF Kit. For each lane, 20 µg of total protein were loaded.

Figure 2. Exosomes prepared using the XCF COMPLETE Kit deliver the expected particle size profile.

NanoSight NTA data for extracellular vesicles isolated from 500 µl of serum (Panel A) and plasma (Panel B) show the expected size profile for exosomes with very little smaller or larger-sized particles.

Figure 3. cfDNA and exosomal DNA isolated using the XCF COMPLETE Kit are of good quality and yield with the expected size.

Agilent Bioanalyzer profiles of the exosomal DNA portion (Panel A) and cfDNA portion (Panel B) of 500 µl of serum processed with the XCF COMPLETE Kit. A notable peak around ~166 bp is seen in both cfDNA and exosomal DNA, which is consistent with cfDNA sizes reported in literature1, and overall DNA quality and yields are sufficient for downstream applications such as qPCR and sequencing.

References

1. Lo YM, et al. Maternal plasma DNA sequencing reveals the genome-wide genetic and mutational profile of the fetus. Sci Transl Med. 2010. 2(61):61ra91. PMID: 21148127.