Cancer Genetics and Epigenetics, 2025, Vol.13, No.3, 106-116 http://medscipublisher.com/index.php/cge 110 In addition, exosome sampling is non-invasive and convenient for multiple monitoring. Doctors can thereby track the changes in the condition, discover residual lesions, and evaluate the treatment effect in a timely manner (Kang et al., 2020; Breakefield et al., 2021; Tusubira et al., 2022). This ability of dynamic monitoring is conducive to formulating more personalized and flexible cancer treatment plans, which is expected to improve the prognosis of patients and reduce unnecessary invasive surgeries (Jafarbeik-Iravani et al., 2017; Song et al., 2020; Cai et al., 2022; Wang et al., 2025). 5 Advances in Exosome Analysis Technology 5.1 Separation techniques: supercentrifugation, nanomaterials, immunoaffinity method Supercentrifugation remains a commonly used method for separating exosomes. It relies on the centrifugal force generated by high-speed rotation to separate exosomes based on their size and density. Although supercentrifugation can better maintain the morphology of exosomes, the operation takes a long time, requires a large sample size, and impurities are prone to mix in during separation, resulting in the aggregation of exosomes and reducing purity (Hsu et al., 2019; Gopal et al., 2021). Precipitation-based methods such as total exosome separation reagents can separate more and purer exosomes, but they may also introduce non-exosome particles. Therefore, they should be carefully selected based on subsequent uses when in use (Burgy et al., 2020; Kumar et al., 2025). Nowadays, new technologies based on nanomaterials and the principle of immunoaffinity can be used to separate exosomes more precisely and efficiently. Microfluidic devices based on nanomaterials, along with aptamer capture technology, feature fast separation speed, high yield, good purity, and can also reduce sample usage and operation time (Demirci et al., 2021; Du et al., 2022). The immunoaffinity method separates exosome surface markers through antibodies or aptamers, but it has the problems of high cost and difficulty in large-scale application. The combination of these advanced methods is promoting the development of exosome isolation technology in clinical practice and research (Hsu et al., 2019; Shen et al., 2023; Hu and Gao, 2025). 5.2 Detection methods: qPCR, NGS, nanosensors, single-molecule analysis Quantitative PCR (qPCR) and next-generation sequencing (NGS) are important methods for detecting nucleic acids in exosomes, which can accurately detect and analyze genetic materials such as micrornas and messenger Rnas in exosomes These methods are highly specific and crucial for discovering disease-related biomarkers, but the prerequisite is to have a reliable exosome isolation and nucleic acid extraction process to ensure the accuracy of the results (Gopal et al., 2021; Yang et al., 2024). New achievements in nanosensor technology and single-molecule analysis have enhanced the ability to detect exosomes. Fluorescence sensors, electrochemical sensors, surface-enhanced Raman scattering (SERS) technology, and microfluidic-based biosensors, etc., can rapidly and sensitively detect proteins and nucleic acids in exosomes without labeling (Figure 2) (Demirci et al., 2021; Hu and Gao, 2025). These techniques can also be combined with signal amplification methods such as hybrid chain reaction or rolling loop amplification to improve detection accuracy and facilitate rapid diagnosis (Huang et al., 2021; Yang et al., 2024). 5.3 Technical challenges and standardization requirements: purity, sensitivity, repeatability Although exosome analysis technology has made significant progress, it still faces many challenges, especially in terms of the purity, sensitivity and repeatability of results in separation and detection. When isolating exosomes by traditional methods, impurities such as protein aggregates and lipoproteins are prone to be mixed in, which brings difficulties to subsequent analysis and the discovery of biomarkers (Burgy et al., 2020; Huang et al., 2021). Moreover, the methods for isolating exosomes in various laboratories are not uniform and there is a lack of standard procedures, resulting in inconsistent experimental results and hindering the transformation of related technologies into clinical applications (Hsu et al., 2019; Gopal et al., 2021; Du et al., 2022). There is an urgent need to formulate standard and reliable operation procedures now to ensure the high purity, high sensitivity and result repeatability of exosome analysis. The development of a new platform integrating microfluidics and biosensors, while unifying the norms of sample processing and data analysis, is crucial for
RkJQdWJsaXNoZXIy MjQ4ODYzNA==