Tandem femto- and nanomolar analysis of two protein biomarkers in plasma on a single mixed antibody monolayer surface using surface plasmon resonance

Suhee Kim, Jeong Won Park, Alastair W. Wark, Sung Hwa Jhung, Hye Jin Lee

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

The multiplexed detection of protein biomarkers in plasma present over a range of clinically relevant concentrations continues to be difficult for surface-based bioaffinity detection platforms such as surface plasmon resonance (SPR). As well as nonspecific adsorption, challenges include quantitative comparison between targets whose concentrations differ by orders of magnitude, regenerating SPR chips after plasma exposure, and the two- or four-channel limitation of many commercial SPR instruments limiting sample throughput. In this article, we explore an approach where two protein biomarkers alpha-1 antitrypsin (AAT) and Tau 381 are detected in tandem within a single SPR channel at micromolar and femtomolar concentrations, respectively. This was achieved by creating a mixed antibody (antiAAT and antiTau) monolayer on the chip surface. After the adsorption of AAT and/or Tau, further specificity was obtained via the adsorption of a DNA aptamer specific to each target. The detection range for each target was controlled via the relative surface density ratio of each antibody type as well as each aptamer concentration. Calibration measurements were performed in both buffer and spiked plasma with the detection of native concentrations of ∼39 fM (Tau) and ∼65 μM (AAT) in a human plasma sample. Finally, tandem measurements of both targets within the same SPR signal channel were demonstrated at these very different concentrations.
LanguageEnglish
Pages12562-12568
Number of pages7
JournalAnalytical Chemistry
Volume89
Issue number22
Early online date25 Oct 2017
DOIs
Publication statusPublished - 21 Nov 2017

Fingerprint

Surface plasmon resonance
Biomarkers
Monolayers
alpha 1-Antitrypsin
Plasmas
Antibodies
Proteins
Adsorption
Nucleotide Aptamers
Plasma (human)
Buffers
Throughput
Calibration

Keywords

  • plasma
  • protein biomarkers
  • surface plasmon resonance

Cite this

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title = "Tandem femto- and nanomolar analysis of two protein biomarkers in plasma on a single mixed antibody monolayer surface using surface plasmon resonance",
abstract = "The multiplexed detection of protein biomarkers in plasma present over a range of clinically relevant concentrations continues to be difficult for surface-based bioaffinity detection platforms such as surface plasmon resonance (SPR). As well as nonspecific adsorption, challenges include quantitative comparison between targets whose concentrations differ by orders of magnitude, regenerating SPR chips after plasma exposure, and the two- or four-channel limitation of many commercial SPR instruments limiting sample throughput. In this article, we explore an approach where two protein biomarkers alpha-1 antitrypsin (AAT) and Tau 381 are detected in tandem within a single SPR channel at micromolar and femtomolar concentrations, respectively. This was achieved by creating a mixed antibody (antiAAT and antiTau) monolayer on the chip surface. After the adsorption of AAT and/or Tau, further specificity was obtained via the adsorption of a DNA aptamer specific to each target. The detection range for each target was controlled via the relative surface density ratio of each antibody type as well as each aptamer concentration. Calibration measurements were performed in both buffer and spiked plasma with the detection of native concentrations of ∼39 fM (Tau) and ∼65 μM (AAT) in a human plasma sample. Finally, tandem measurements of both targets within the same SPR signal channel were demonstrated at these very different concentrations.",
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Tandem femto- and nanomolar analysis of two protein biomarkers in plasma on a single mixed antibody monolayer surface using surface plasmon resonance. / Kim, Suhee; Park, Jeong Won; Wark, Alastair W.; Jhung, Sung Hwa; Lee, Hye Jin.

In: Analytical Chemistry, Vol. 89, No. 22, 21.11.2017, p. 12562-12568.

Research output: Contribution to journalArticle

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T1 - Tandem femto- and nanomolar analysis of two protein biomarkers in plasma on a single mixed antibody monolayer surface using surface plasmon resonance

AU - Kim, Suhee

AU - Park, Jeong Won

AU - Wark, Alastair W.

AU - Jhung, Sung Hwa

AU - Lee, Hye Jin

N1 - This document is the Accepted Manuscript version of a Published Work that appeared in final form in Analytical Chemistry, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://doi.org/10.1021/acs.analchem.7b03837.

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Y1 - 2017/11/21

N2 - The multiplexed detection of protein biomarkers in plasma present over a range of clinically relevant concentrations continues to be difficult for surface-based bioaffinity detection platforms such as surface plasmon resonance (SPR). As well as nonspecific adsorption, challenges include quantitative comparison between targets whose concentrations differ by orders of magnitude, regenerating SPR chips after plasma exposure, and the two- or four-channel limitation of many commercial SPR instruments limiting sample throughput. In this article, we explore an approach where two protein biomarkers alpha-1 antitrypsin (AAT) and Tau 381 are detected in tandem within a single SPR channel at micromolar and femtomolar concentrations, respectively. This was achieved by creating a mixed antibody (antiAAT and antiTau) monolayer on the chip surface. After the adsorption of AAT and/or Tau, further specificity was obtained via the adsorption of a DNA aptamer specific to each target. The detection range for each target was controlled via the relative surface density ratio of each antibody type as well as each aptamer concentration. Calibration measurements were performed in both buffer and spiked plasma with the detection of native concentrations of ∼39 fM (Tau) and ∼65 μM (AAT) in a human plasma sample. Finally, tandem measurements of both targets within the same SPR signal channel were demonstrated at these very different concentrations.

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