Dried blood sample analysis by antibody array across the total testing process

Whittaker, Kelly; Mao, Yu; Lin, Yongping; Zhang, Huihua; Zhu, Siwei; Peck, Hannah E.; Huang, Ruo‐Pan

doi:10.1038/s41598-021-99911-8

Cited by 8 publications

(7 citation statements)

References 56 publications

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“…We performed a one-way ANOVA to compare the effect of five different hematocrit values on the quantitation of transferrin receptor protein from pDPS cards, which revealed that there was not a statistically significant difference between mean transferrin receptor protein concentrations for whole blood samples with varying hematocrit values ( F = [1.893] and p = 0.15). The overall yield of analytes eluted from TFN can be improved by introducing low concentrations of surfactants into the elution buffer to encourage the resolubilization of proteins following the drying step (vide infra) . However, maintaining a consistent concentration of this soluble analyte provided confidence in the volume metering capabilities of this device.…”

Section: Resultsmentioning

confidence: 99%

“…The overall yield of analytes eluted from TFN can be improved by introducing low concentrations of surfactants into the elution buffer to encourage the resolubilization of proteins following the drying step (vide infra). 44 However, maintaining a consistent concentration of this soluble analyte provided confidence in the volume metering capabilities of this device. Additionally, reproducibly producing a discrete volume of liquid plasma� independent of the hematocrit�can enable quantitative analysis to aid in a clinical diagnosis rather than simply detecting the presence of an analyte.…”

Section: ■ Results and Discussion Pdps Cards Reproducibly Fill Indepe...mentioning

confidence: 99%

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Direct Processing and Storage of Cell-Free Plasma Using Dried Plasma Spot Cards

et al. 2022

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Plasma separation cards represent a viable approach for expanding testing capabilities away from clinical settings by generating cell-free plasma with minimal user intervention. These devices typically comprise a basic structure of the plasma separation membrane, unconstrained porous collection pad, and utilize either (i) lateral or (ii) vertical fluidic pathways for separating plasma. Unfortunately, these configurations are highly susceptible to (i) inconsistent sampling volume due to differences in the patient hematocrit or (ii) severe contamination due to leakage of red blood cells or release of hemoglobin (i.e., hemolysis). Herein, we combine the enhanced sampling of our previously reported patterned dried blood spot cards with an assembly of porous separation materials to produce a patterned dried plasma spot card for direct processing and storage of cell-free plasma. Linking both vertical separation and lateral distribution of plasma yields discrete plasma collection zones that are spatially protected from potential contamination due to hemolysis and an inlet zone enriched with blood cells for additional testing. We evaluate the versatility of this card by quantitation of three classes of analytes and techniques including (i) the soluble transferrin receptor by enzyme-linked immunosorbent assay, (ii) potassium by inductively coupled plasma atomic emission spectroscopy, and (iii) 18S rRNA by reverse transcriptase quantitative polymerase chain reaction. We achieve quantitative recovery of each class of analyte with no statistically significant difference between dried and liquid reference samples. We anticipate that this sampling approach can be applied broadly to improve access to critical blood testing in resource-limited settings or at the point-of-care.

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Section: Resultsmentioning

confidence: 99%

Section: ■ Results and Discussion Pdps Cards Reproducibly Fill Indepe...mentioning

confidence: 99%

Direct Processing and Storage of Cell-Free Plasma Using Dried Plasma Spot Cards

et al. 2022

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show abstract

“…The overall yield of analyte eluted from TFN can be improved by introducing low concentrations of surfactants into the elution buffer to encourage the re-solubilization of proteins following the drying step (vide infra). 43 However, maintaining a consistent concentration of this soluble analyte provided confidence in the volume metering capabilities of this device. Additionally, reproducibly producing a discrete volume of liquid plasma-independently of the hematocrit-can enable quantitative analysis to aid in a clinical diagnosis rather than simply detecting the presence of an analyte.…”

Section: Pdps Cards Reproducibly Fill Independently Of Hematocrit Valuementioning

confidence: 99%

Direct Processing and Storage of Cell-free Plasma using Dried Plasma Spot Cards

Baillargeon

Morbioli

Brooks

et al. 2022

Preprint

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Plasma separation cards represent a viable approach for expanding testing capabilities away from clinical settings by generating cell-free plasma with minimal user intervention. These devices typically comprise a basic structure of plasma separation membrane (PSM), unconstrained porous collection pad, and utilize either (i) lateral or (ii) vertical fluidic pathways for separating plasma. Unfortunately, these configurations are highly susceptible to (i) inconsistent sampling volume due to differences in patient hematocrit or (ii) severe contamination due to leakage of red blood cells (RBCs) or release of hemoglobin (i.e., hemolysis). Herein, we combine the enhanced sampling of our previously reported patterned dried blood spot (pDBS) cards with an assembly of porous separation materials to produce a patterned dried plasma spot (pDPS) card for direct processing and storage of cell-free plasma. Linking both vertical separation and lateral distribution of plasma yields discrete plasma collection zones that are spatially protected from potential contamination due to hemolysis and an inlet zone enriched with blood cells for additional testing. We evaluate the versatility of this card by quantitation of three classes of analytes and techniques including: (i) soluble transferrin receptor by enzyme-linked immunosorbent assay (ELISA), (ii) potassium by inductively coupled plasma atomic emission spectroscopy (ICP-AES), and (iii) 18S rRNA by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR). We achieve quantitative recovery of each class of analyte with no statistically significant difference between dried and liquid reference samples. We anticipate that this sampling approach can be applied broadly to improve access to critical blood testing in resource limited settings or at the point of care.

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“… 25 , 26 Despite a number of issues related to DBS sampling, such as the impact of hematocrit on accurate quantification, 27 , 28 devices for accurate volumetric collection 29 have proven to overcome most issues associated with conventional DBS collection. 30 − 32 …”

Section: Introductionmentioning

confidence: 99%

“…Dried blood spot (DBS) sampling is a patient-centric sampling method, used to collect, ship, and store blood, that has the potential to alleviate some of the problems associated with the preanalytical phase. However, the capability to accurately quantify low-abundant biomarkers relies on the quality of the dried sample, the ability to quantify from it (through volume or intrinsic markers in the sample), the efficiency of recovery upon elution, and the sensitivity of the assay downstream since the elution procedure may lead to a dilution of 5–10 times or even more. , Despite a number of issues related to DBS sampling, such as the impact of hematocrit on accurate quantification, , devices for accurate volumetric collection have proven to overcome most issues associated with conventional DBS collection. − …”

Section: Introductionmentioning

confidence: 99%

Microfluidic Device for Patient-Centric Multiplexed Assays with Readout in Centralized Laboratories

et al. 2022

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Patient-centric sampling strategies, where the patient performs self-sampling and ships the sample to a centralized laboratory for readout, are on the verge of widespread adaptation. However, the key to a successful patient-centric workflow is user-friendliness, with few noncritical user interactions, and simple, ideally biohazard-free shipment. Here, we present a capillary-driven microfluidic device designed to perform the critical biomarker capturing step of a multiplexed immunoassay at the time of sample collection. On-chip sample drying enables biohazard-free shipment and allows us to make use of advanced analytics of specialized laboratories that offer the needed analytical sensitivity, reliability, and affordability. Using C-Reactive Protein, MCP1, S100B, IGFBP1, and IL6 as model blood biomarkers, we demonstrate the multiplexing capability and applicability of the device to a patient-centric workflow. The presented quantification of a biomarker panel opens up new possibilities for e-doctor and e-health applications.

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Dried blood sample analysis by antibody array across the total testing process

Cited by 8 publications

References 56 publications

Direct Processing and Storage of Cell-Free Plasma Using Dried Plasma Spot Cards

Direct Processing and Storage of Cell-Free Plasma Using Dried Plasma Spot Cards

Direct Processing and Storage of Cell-free Plasma using Dried Plasma Spot Cards

Microfluidic Device for Patient-Centric Multiplexed Assays with Readout in Centralized Laboratories

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