The use of hydrogel microparticles to sequester and concentrate bacterial antigens in a urine test for Lyme disease

Temple A. Douglas, Davide Tamburro, Claudia Fredolini, Benjamin H. Espina, Benjamin S. Lepene, Leopold Ilag, Virginia Espina, Emanuel F. Petricoin, Lance A. Liotta, Alessandra Luchini*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

43 Scopus citations

Abstract

Hydrogel biomarker capturing microparticles were evaluated as a biomaterial to amplify the sensitivity of urine testing for infectious disease proteins. Lyme disease is a bacterial infection transmitted by ticks. Early diagnosis and prompt treatment of Lyme disease reduces complications including arthritis and cardiac involvement. While a urine test is highly desirable for Lyme disease screening, this has been difficult to accomplish because the antigen is present at extremely low concentrations, below the detection limit of clinical immunoassays. N-isopropylacrylamide (NIPAm) - acrylic acid (AAc) microparticles were covalently functionalized with amine containing dyes via amidation of carboxylic groups present in the microparticles. The dyes act as affinity baits towards protein analytes in solution. NIPAm/AAc microparticles functionalized with acid black 48 (AB48) mixed with human urine, achieved close to one hundred percent capture and 100 percent extraction yield of the target antigen. In urine, microparticles sequestered and concentrated Lyme disease antigens 100 fold, compared to the absence of microparticles, achieving an immunoassay detection sensitivity of 700 pg/mL in 10 mL urine. Antigen present in a single infected tick could be readily detected following microparticle sequestration. Hydrogel microparticles functionalized with high affinity baits can dramatically increase the sensitivity of urinary antigen tests for infectious diseases such as Lyme disease. These findings justify controlled clinical studies evaluating the sensitivity and precision of Lyme antigen testing in urine.

Original languageEnglish
Pages (from-to)1157-1166
Number of pages10
JournalBiomaterials
Volume32
Issue number4
DOIs
StatePublished - Feb 2011
Externally publishedYes

Keywords

  • Affinity
  • Bacteria
  • Hydrogel
  • Microparticle
  • Protein

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