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Development of a new biochip array for APOE4 classification from plasma samples using immunoassay-based methods

  • Sigrun Badrnya , Tara Doherty , Ciaran Richardson , Robert I. McConnell , John V. Lamont , Michael Veitinger , Stephen P. FitzGerald , Maria Zellner and Ellen Umlauf ORCID logo EMAIL logo
Published/Copyright: December 7, 2017
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Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 56 Issue 5

Abstract

Background:

Apolipoprotein E (APOE) is a key player in lipid transport and metabolism and exists in three common isoforms: APOE2, APOE3 and APOE4. The presence of the E4 allelic variant is recognized as a major genetic risk factor for dementia and other chronic (neuro)degenerative diseases. The availability of a validated assay for rapid and reliable APOE4 classification is therefore advantageous.

Methods:

Biochip array technology (BAT) was successfully applied to identify directly the APOE4 status from plasma within 3 h, through simultaneous immunoassay-based detection of both specific APOE4 and total APOE levels.

Results:

Samples (n=432) were first genotyped by polymerase chain reaction (PCR), and thereafter, using BAT, the corresponding plasma was identified as null, heterozygous or homozygous for the E4 allele by calculating the ratio of APOE4 to total APOE protein. Two centers based in Austria and Ireland correctly classified 170 and 262 samples, respectively, and achieved 100% sensitivity and specificity.

Conclusions:

This chemiluminescent biochip-based sandwich immunoarray provides a novel platform to detect rapidly and accurately an individual’s APOE4 status directly from plasma. The E4 genotype of individuals has been shown previously to affect presymptomatic risk, prognosis and treatment response for a variety of diseases, including Alzheimer’s disease. The biochip’s potential for being incorporated in quantitative protein biomarker arrays capable of analyzing disease stages makes it a superior alternative to PCR-based APOE genotyping and may deliver additional protein-specific information on a variety of diseases in the future.

Keywords: APOE4 phenotyping; apolipoprotein E4; biochip; blood test; cardiovascular disease; dementia; genetic risk factor; late-onset Alzheimer’s disease; multiarray; neurodegeneration
Corresponding author: Dr. Ellen Umlauf, Centre of Physiology and Pharmacology, Institute of Physiology, Medical University of Vienna, Schwarzspanierstr. 17, 1090 Vienna, Austria

Acknowledgments

Egon Ogris and Marko Roblek developed the anti-APOE4 antibody. We thank Aner Gurvitz for critically reading and editing the manuscript.

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: This work was supported by the FP7 framework program of the European Commission (Marie-Curie Actions, funder ID: 10.13039/100011264, grant no. FP7PEOPLE2011IAPP286337).

  3. Employment or leadership: None declared.

  4. Honorarium: None declared

  5. Competing interests: The funding organization played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

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Received: 2017-7-14
Accepted: 2017-10-31
Published Online: 2017-12-7
Published in Print: 2018-4-25

©2018 Walter de Gruyter GmbH, Berlin/Boston

Articles in the same Issue

  1. Frontmatter
  2. Editorials
  3. Scientific publishing in the “predatory” era
  4. The influence of age and other biological variables on the estimation of reference limits of cardiac troponin T
  5. Reviews
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  9. Practical recommendations for managing hemolyzed samples in clinical chemistry testing
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  28. Detection of EGFR, KRAS and BRAF mutations in metastatic cells from cerebrospinal fluid
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  31. Letters to the Editor
  32. Rare inclusion bodies within monocytes at accelerated phase of Chediak-Higashi syndrome
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  35. Performance evaluation of a new automated fourth-generation HIV Ag/Ab combination chemiluminescence immunoassay
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https://doi.org/10.1515/cclm-2017-0618
Keywords for this article
APOE4 phenotyping; apolipoprotein E4; biochip; blood test; cardiovascular disease; dementia; genetic risk factor; late-onset Alzheimer’s disease; multiarray; neurodegeneration

Articles in the same Issue

  1. Frontmatter
  2. Editorials
  3. Scientific publishing in the “predatory” era
  4. The influence of age and other biological variables on the estimation of reference limits of cardiac troponin T
  5. Reviews
  6. Prognostic and predictive value of EGFR and EGFR-ligands in blood of breast cancer patients: a systematic review
  7. GSTP1 methylation in cancer: a liquid biopsy biomarker?
  8. Opinion Paper
  9. Practical recommendations for managing hemolyzed samples in clinical chemistry testing
  10. Genetics and Molecular Diagnostics
  11. Non-invasive prenatal diagnosis of paternally inherited disorders from maternal plasma: detection of NF1 and CFTR mutations using droplet digital PCR
  12. Circulating miR-21, miR-210 and miR-146a as potential biomarkers to differentiate acute tubular necrosis from hepatorenal syndrome in patients with liver cirrhosis: a pilot study
  13. Pleiotropy of ABO gene: correlation of rs644234 with E-selectin and lipid levels
  14. General Clinical Chemistry and Laboratory Medicine
  15. Three-year customer satisfaction survey in laboratory medicine in a Chinese university hospital
  16. Measurement of sirolimus concentrations in human blood using an automated electrochemiluminescence immunoassay (ECLIA): a multicenter evaluation
  17. Precision, accuracy, cross reactivity and comparability of serum indices measurement on Abbott Architect c8000, Beckman Coulter AU5800 and Roche Cobas 6000 c501 clinical chemistry analyzers
  18. Commutability of control materials for external quality assessment of serum apolipoprotein A-I measurement
  19. Development of a new biochip array for APOE4 classification from plasma samples using immunoassay-based methods
  20. Validation of a high-performance liquid chromatography method for thiopurine S-methyltransferase activity in whole blood using 6-mercaptopurine as substrate
  21. Increased serum concentrations of soluble ST2 are associated with pulmonary complications and mortality in polytraumatized patients
  22. Reference Values and Biological Variations
  23. Determination of age- and sex-specific 99th percentiles for high-sensitive troponin T from patients: an analytical imprecision- and partitioning-based approach
  24. Effect of preanalytical and analytical variables on the clinical utility of mean platelet volume
  25. Serum prolactin levels across pregnancy and the establishment of reference intervals
  26. Gender-partitioned patient medians of serum albumin requested by general practitioners for the assessment of analytical stability
  27. Cancer Diagnostics
  28. Detection of EGFR, KRAS and BRAF mutations in metastatic cells from cerebrospinal fluid
  29. Cardiovascular Diseases
  30. No additional value of conventional and high-sensitivity cardiac troponin over clinical scoring systems in the differential diagnosis of type 1 vs. type 2 myocardial infarction
  31. Letters to the Editor
  32. Rare inclusion bodies within monocytes at accelerated phase of Chediak-Higashi syndrome
  33. A specific abnormal scattergram of peripheral blood leukocytes that may suggest hairy cell leukemia
  34. Spuriously low lymphocyte count associated with pseudoerythroblastemia in a patient with chronic lymphocytic leukemia treated with ibrutinib
  35. Performance evaluation of a new automated fourth-generation HIV Ag/Ab combination chemiluminescence immunoassay
  36. False increase of glycated hemoglobin due to aspirin interference in Tosoh G8 analyzer
  37. Assessment of in vitro stability: a call for harmonization across studies
  38. Comparison between blood gas analyzer and central laboratory analyzer for the determination of electrolytes in patients with acute respiratory acidosis
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