An examination of linking and blocking procedures for use in deflection cantilever array-based protein detection

• Author / Creator

  van den Hurk, Remko

• In this project common linking and blocking procedures were examined for use in multiplexed deflection cantilever array-based detection of proteins. A human interferon gamma (INF-γ) enzyme-linked immunosorbent assay (ELISA) kit was used to evaluate the effectiveness of the linking and blocking procedures using fluorescence and ELISA assays. The most effective linking and blocking procedures were then implemented on arrays of deflection cantilevers. Separating the active and reference cantilever signals proved to be challenging due to varying deflection in buffer solution. Background subtractions were implemented which reduced the nonspecific buffer deflection. Following the background subtractions 16 out of 52 experiments produced one or more data sets with a clearly defined separation between all the active and reference signals. The most successful linkers, reference cantilever blockers and cantilever backside blockers were Prolinker B, glutaradehyde and EDC/Sulfo-NHS; canine capture antibody and INF-γ; and a thermal PEG-silanization procedure and PEG-thiol, respectively.

• Subjects / Keywords

  • Cantilever
  • Linking
  • Biosensors
  • Proteins
  • MEMS
  • Detection
  • Blocking
  • ELISA
  • Fluorescence
  • Array

• Graduation date

  Fall 2011

• Type of Item

  Thesis

• Degree

  Master of Science

• DOI

  https://doi.org/10.7939/R3V701

• License

  This thesis is made available by the University of Alberta Libraries with permission of the copyright owner solely for non-commercial purposes. This thesis, or any portion thereof, may not otherwise be copied or reproduced without the written consent of the copyright owner, except to the extent permitted by Canadian copyright law.