TY - JOUR
T1 - The effects of flow type on aptamer capture in differential mobility cytometry cell separations
AU - Liu, Yan
AU - Bae, Se Won
AU - Wang, Kelong
AU - Hong, Jong In
AU - Zhu, Zhi
AU - Tan, Weihong
AU - Pappas, Dimitri
PY - 2010/7
Y1 - 2010/7
N2 - In this work, differential mobility cytometry (DMC) was used to monitor cell separation based on aptamer recognition for target cells. In this device, open-tubular capillaries coated with Sgc8 aptamers were used as affinity chromatography columns for separation. After cells were injected into the columns, oscillating flow was generated to allow for long-term cell adhesion studies. This process was monitored by optical microscopy, and differential imaging was used to analyze the cells as they adhered to the affinity surface. We investigated the capture time, capture efficiency, purity of target and control cells, as well as the reusability of the affinity columns. Capture time for both CCRF-CEM cells and Jurkat T cells was 0.4 ± 0.2 s, which demonstrated the high separation affinity between aptamers and target cells. The capture efficiency for CCRF-CEM cells was 95% and purity was 99% in a cell mixture. With the advantage of both high cell capture efficiency and purity, DMC combined with aptamer-based separation emerges as a powerful tool for rare cell enrichment. In addition, aptamer-based DMC channels were found to be more robust than antibody based channels with respect to reuse of the separation device.
AB - In this work, differential mobility cytometry (DMC) was used to monitor cell separation based on aptamer recognition for target cells. In this device, open-tubular capillaries coated with Sgc8 aptamers were used as affinity chromatography columns for separation. After cells were injected into the columns, oscillating flow was generated to allow for long-term cell adhesion studies. This process was monitored by optical microscopy, and differential imaging was used to analyze the cells as they adhered to the affinity surface. We investigated the capture time, capture efficiency, purity of target and control cells, as well as the reusability of the affinity columns. Capture time for both CCRF-CEM cells and Jurkat T cells was 0.4 ± 0.2 s, which demonstrated the high separation affinity between aptamers and target cells. The capture efficiency for CCRF-CEM cells was 95% and purity was 99% in a cell mixture. With the advantage of both high cell capture efficiency and purity, DMC combined with aptamer-based separation emerges as a powerful tool for rare cell enrichment. In addition, aptamer-based DMC channels were found to be more robust than antibody based channels with respect to reuse of the separation device.
KW - Aptamer
KW - Cell capture time
KW - Cell separation
KW - Differential mobility cytometry
UR - http://www.scopus.com/inward/record.url?scp=77954143573&partnerID=8YFLogxK
U2 - 10.1016/j.aca.2010.05.017
DO - 10.1016/j.aca.2010.05.017
M3 - Article
AN - SCOPUS:77954143573
SN - 0003-2670
VL - 673
SP - 95
EP - 100
JO - Analytica Chimica Acta
JF - Analytica Chimica Acta
IS - 1
ER -