Development of a Binding-Induced DNA Assembly Assay for Carbohydrate Antigen 19-9

• Author / Creator

  Wagner, Megan K.

• Binding-induced DNA assembly (BINDA) was initially developed for protein detection. To demonstrate the homogeneous detection of carbohydrates using this principle, a BINDA assay for the pancreatic cancer marker carbohydrate antigen 19-9 (CA 19-9) was developed. The assay uses two DNA- antibody conjugates as probes, with the DNA oligonucleotides designed to have a complementary base-pairing sequence. Binding to CA 19-9 brings the probes into close proximity, allowing the complementary regions to hybridize. Enzymatic ligation joins the probe oligonucleotides and amplification and detection of the ligation product by real-time PCR is used to quantify CA 19-9. The principle was first demonstrated using streptavidin as a model target before developing and optimizing the BINDA assay for CA 19-9. CA 19-9 could be detected in both buffer and human serum. This study is the first description of the use of BINDA for carbohydrate analysis.

• Subjects / Keywords

  • Real-time PCR
  • Cancer marker detection
  • Pancreatic cancer
  • DNA assembly
  • CA 19-9
  • Assay
  • Carbohydrate antigen 19-9

• Graduation date

  Spring 2013

• Type of Item

  Thesis

• Degree

  Master of Science

• DOI

  https://doi.org/10.7939/R39S1KS28

• 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.