title: Analytical evaluation for somatic mutation detection in circulating tumor cells isolated using a lateral magnetophoretic microseparator.
abstract: CTCs are currently in the spotlight because provide comprehensive genetic information that enables monitoring of the evolution of cancer and selection of appropriate therapeutic strategies that cannot be obtained from a single-site tumor biopsy. Despite their importance, current techniques for isolating CTCs are limited in terms of their ability to yield high-quality CTCs from peripheral blood for use in profiling cancer genetic mutations by DNA sequencing technologies. This paper introduces a lateral magnetophoretic microseparator (the 'CTC-μChip') for isolating highly pure CTCs from blood, which facilitates the detection of somatic mutations in isolated CTCs. To isolate CTCs from peripheral blood, nucleated cells were first prepared by red blood cell lysis. Then, CTCs were isolated from nucleated cells within 30 min using the CTC-μChip. Analytical evaluation using 5 mL blood samples spiked with 5-50 MCF7 breast cancer cells demonstrated that the average recovery rate of the CTC-μChip was 99.08 %. The average number of residual white blood cells (WBCs) in isolated samples was 53, meaning that the WBC depletion rate is 472,000-fold (5.67 log), assuming that blood contains 5 × 10(6) WBCs per milliliter. The isolated MCF7 cells had a purity of 6.9 - 67.9 %, depending on the spiked MCF7 concentration. Using next-generation sequencing technology, heterozygous somatic mutations (PIK3CA and APC) of MCF7 cells were evaluated in the isolated samples. The results showed that somatic mutations could be detected in as few as two MCF7 cells per milliliter of blood, indicating that the CTC-μChip facilitates the detection of somatic variants in CTCs.
authors.affiliation: Department of Nano Science and Engineering Center for Nano Manufacturing, Inje University, 607 Obang-dong, Gimhae, Gyongnam, 621-749, Republic of Korea.
authors.forename: Hyungseok
authors.initials: H
authors.lastname: Cho
authors.affiliation: Department of Nano Science and Engineering Center for Nano Manufacturing, Inje University, 607 Obang-dong, Gimhae, Gyongnam, 621-749, Republic of Korea.
authors.forename: Jinho
authors.initials: J
authors.lastname: Kim
authors.affiliation: Department of Nano Science and Engineering Center for Nano Manufacturing, Inje University, 607 Obang-dong, Gimhae, Gyongnam, 621-749, Republic of Korea.
authors.forename: Song-I
authors.initials: SI
authors.lastname: Han
authors.affiliation: Department of Nano Science and Engineering Center for Nano Manufacturing, Inje University, 607 Obang-dong, Gimhae, Gyongnam, 621-749, Republic of Korea. mems@inje.ac.kr.
authors.forename: Ki-Ho
authors.initials: KH
authors.lastname: Han
mesh_terms: D001943:Breast Neoplasms / Q000473:pathology; D004252:DNA Mutational Analysis; D006801:Humans; D018189:Immunomagnetic Separation / Q000295:instrumentation*; D007963:Leukocytes, Mononuclear / Q000166:cytology; D061986:MCF-7 Cells; D009360:Neoplastic Cells, Circulating / Q000378:metabolism* / Q000473:pathology*; D013194:Staining and Labeling
keywords: CTC-μChip; Cancer; Circulating tumor cells; Next-generation sequencing; Somatic mutation