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Microfluidic Immunoaffinity Basophil Activation Test for Point-of-Care Allergy Diagnosis
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Nano Biotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab. Karolinska Inst, Dept Clin Sci & Educ, Stockholm, Sweden; Soder Sjukhuset, Stockholm, Sweden .
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Nano Biotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab. Karolinska Inst, Dept Clin Sci & Educ, Stockholm, Sweden; Soder Sjukhuset, Stockholm, Sweden.
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Nano Biotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.ORCID iD: 0000-0001-5199-0663
Karolinska Inst, Dept Clin Sci & Educ, Stockholm, Sweden.;Soder Sjukhuset, Stockholm, Sweden..
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2019 (English)In: Journal of Applied Laboratory Medicine (JALM), ISSN 2475-7241, Vol. 4, no 2, p. 152-163Article in journal (Refereed) Published
Abstract [en]

Background: The flow cytometry-based basophil activation test (BAT) is used for the diagnosis of allergic response. However, flow cytometry is time-consuming, requiring skilled personnel and cumbersome processing, which has limited its use in the clinic. Here, we introduce a novel microfluidic-based immunoaffinity BAT (miBAT) method. Methods: The microfluidic device, coated with anti-CD203c, was designed to capture basophils directly from whole blood. The captured basophils are activated by anti-FceRI antibody followed by optical detection of CD63 expression (degranulation marker). The device was first characterized using a basophil cell line followed by whole blood experiments. Weevaluated the device with ex vivo stimulation of basophils in whole blood from healthy controls and patients with allergies and compared it with flow cytometry. Results: The microfluidic device was capable of capturing basophils directly from whole blood followed by in vitro activation and quantification of CD63 expression. CD63 expression was significantly higher (P = 0.0002) in on-chip activated basophils compared with nonactivated cells. The difference in CD63 expression on anti-FceRI-activated captured basophils in microfluidic chip was significantly higher (P = 0.03) in patients with allergies compared with healthy controls, and the results were comparable with flow cytometry analysis (P = 0.04). Furthermore, there was no significant difference of CD63% expression in anti-FceRI-activated captured basophils in microfluidic chip compared with flow cytometry. Conclusions: We report on the miBAT. This device is capable of isolating basophils directly from whole blood for on-chip activation and detection. The new miBAT method awaits validation in larger patient populations to assess performance in diagnosis and monitoring of patients with allergies at the point of care.

Place, publisher, year, edition, pages
American Association for Clinical Chemistry , 2019. Vol. 4, no 2, p. 152-163
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Clinical Medicine
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URN: urn:nbn:se:kth:diva-266201DOI: 10.1373/jalm.2018.026641ISI: 000501139100004PubMedID: 31639660OAI: oai:DiVA.org:kth-266201DiVA, id: diva2:1383976
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QC 20200109

Available from: 2020-01-09 Created: 2020-01-09 Last updated: 2020-01-09Bibliographically approved

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Aljadi, ZenibRamachandraiah, HarishaRussom, Aman

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