Microfluidic Chip based direct triple antibody immunoassay for monitoring patient comparative response to leukemia treat
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Microfluidic Chip based direct triple antibody immunoassay for monitoring patient comparative response to leukemia treatment Kutay İçöz 1,2
&
Ünal Akar 2 & Ekrem Ünal 3
# Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract We report a time and cost-efficient microfluidic chip for screening the leukemia cells having three specific antigens. In this method, the target blast cells are double sorted with immunomagnetic beads and captured by the 3rd antibody immobilized on the gold surface in a microfluidic chip. The captured blast cells in the chip were imaged using a bright-field optical microscope and images were analyzed to quantify the cells. First sorting was performed with nano size immunomagnetic beads and followed by 2nd sorting where micron size immunomagnetic beads were used. The low-cost microfluidic platform is made of PMMA and glass including micro size gold pads. The developed microfluidic platform was optimized with cultured B type lymphoblast cells and tested with the samples of leukemia patients. The 8 bone marrow samples of 4 leukemia patients on the initial diagnosis and on the 15th day after the start of the chemotherapy treatment were tested both with the developed microfluidic platform and the flow cytometry. A 99% statistical agreement between the two methods shows that the microfluidic chip is able to monitor the decrease in the number of blast cells due to the chemotherapy. The experiments with the patient samples demonstrate that the developed system can perform relative measurements and have a potential to monitor the patient response to the applied therapy and to enable personalized dose adjustment. Keywords Magnetic micro/nano particles . Immunoassay . Biochip . Leukemia . Direct triple antibody . Microfluidic -based monitoring; comparative response
1 Introduction Acute lymphoblastic leukemia (ALL) is the most common type of cancer among children in the United States (Siegel et al., 2017). ALL is divided into two subtypes; B precursor (B-ALL), and T precursor (T-ALL) according to the origination. 85% of the children with ALL arise from B cells, whereas %15 of them originates from T cells (Chiaretti et al., 2014).
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10544-020-00503-6) contains supplementary material, which is available to authorized users. * Kutay İçöz [email protected] 1
BioMINDS (Bio Micro/Nano Devices and Sensors) Lab, Department of Electrical and Electronics Engineering, Abdullah Gül University, 38080 Kayseri, Turkey
2
Bioengineering Department, Abdullah Gül University, 38080 Kayseri, Turkey
3
Pediatric Hematology Department, Erciyes University, 38030 Kayseri, Turkey
B-ALL has four main stages based on the maturation of cells; pro-B, common B, pre-B and immature B. The gene expression and cell surface markers at stages display significant heterogeneity. For example, CD19 is a membrane antigen and highly expressed in all stages of ALL, whereas the presence of CD10 antigen defines common ALL
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