Advanced

Optimization of the Small Glycan Presentation for Binding a Tumor-Associated Antibody : Application to the Construction of an Ultrasensitive Glycan Biosensor

Kveton, Filip; Blšáková, Anna; Hushegyi, Andras; Damborsky, Pavel; Blixt, Ola; Jansson, Bo LU and Tkac, Jan (2017) In Langmuir 33(11). p.2709-2716
Abstract

The main aim of the study was to optimize the interfacial presentation of a small antigen-a Tn antigen (N-acetylgalactosamine)-for binding to its analyte anti-Tn antibody. Three different methods for the interfacial display of a small glycan are compared here, including two methods based on the immobilization of the Tn antigen on a mixed self-assembled monolayer (SAM) (2D biosensor) and the third one utilizing a layer of a human serum albumin (HSA) for the immobilization of a glycan forming a 3D interface. Results showed that the 3D interface with the immobilized Tn antigen is the most effective bioreceptive surface for binding its analyte. The 3D impedimetric glycan biosensor exhibited a limit of detection of 1.4 aM, a wide linear... (More)

The main aim of the study was to optimize the interfacial presentation of a small antigen-a Tn antigen (N-acetylgalactosamine)-for binding to its analyte anti-Tn antibody. Three different methods for the interfacial display of a small glycan are compared here, including two methods based on the immobilization of the Tn antigen on a mixed self-assembled monolayer (SAM) (2D biosensor) and the third one utilizing a layer of a human serum albumin (HSA) for the immobilization of a glycan forming a 3D interface. Results showed that the 3D interface with the immobilized Tn antigen is the most effective bioreceptive surface for binding its analyte. The 3D impedimetric glycan biosensor exhibited a limit of detection of 1.4 aM, a wide linear range (6 orders of magnitude), and high assay reproducibility with an average relative standard deviation of 4%. The buildup of an interface was optimized using various techniques with the visualization of the glycans on the biosensor surface by atomic force microscopy. The study showed that the 3D biosensor is not only the most sensitive compared to other two biosensor platforms but that the Tn antigen on the 3D biosensor surface is more accessible for antibody binding with better kinetics of binding (t50% = 137 s, t50% = the time needed to attain 50% of a steady-state signal) compared to the 2D biosensor configuration with t50% = 354 s. The 3D glycan biosensor was finally applied for the analysis of a human serum sample spiked with an analyte.

(Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Langmuir
volume
33
issue
11
pages
8 pages
publisher
The American Chemical Society
external identifiers
  • scopus:85016159303
  • wos:000397478200003
ISSN
0743-7463
DOI
10.1021/acs.langmuir.6b04021
language
English
LU publication?
yes
id
1515fd98-2c7a-4384-bc52-69f056d37542
date added to LUP
2017-04-05 12:02:44
date last changed
2018-01-07 11:57:56
@article{1515fd98-2c7a-4384-bc52-69f056d37542,
  abstract     = {<p>The main aim of the study was to optimize the interfacial presentation of a small antigen-a Tn antigen (N-acetylgalactosamine)-for binding to its analyte anti-Tn antibody. Three different methods for the interfacial display of a small glycan are compared here, including two methods based on the immobilization of the Tn antigen on a mixed self-assembled monolayer (SAM) (2D biosensor) and the third one utilizing a layer of a human serum albumin (HSA) for the immobilization of a glycan forming a 3D interface. Results showed that the 3D interface with the immobilized Tn antigen is the most effective bioreceptive surface for binding its analyte. The 3D impedimetric glycan biosensor exhibited a limit of detection of 1.4 aM, a wide linear range (6 orders of magnitude), and high assay reproducibility with an average relative standard deviation of 4%. The buildup of an interface was optimized using various techniques with the visualization of the glycans on the biosensor surface by atomic force microscopy. The study showed that the 3D biosensor is not only the most sensitive compared to other two biosensor platforms but that the Tn antigen on the 3D biosensor surface is more accessible for antibody binding with better kinetics of binding (t<sub>50%</sub> = 137 s, t<sub>50%</sub> = the time needed to attain 50% of a steady-state signal) compared to the 2D biosensor configuration with t<sub>50%</sub> = 354 s. The 3D glycan biosensor was finally applied for the analysis of a human serum sample spiked with an analyte.</p>},
  author       = {Kveton, Filip and Blšáková, Anna and Hushegyi, Andras and Damborsky, Pavel and Blixt, Ola and Jansson, Bo and Tkac, Jan},
  issn         = {0743-7463},
  language     = {eng},
  month        = {03},
  number       = {11},
  pages        = {2709--2716},
  publisher    = {The American Chemical Society},
  series       = {Langmuir},
  title        = {Optimization of the Small Glycan Presentation for Binding a Tumor-Associated Antibody : Application to the Construction of an Ultrasensitive Glycan Biosensor},
  url          = {http://dx.doi.org/10.1021/acs.langmuir.6b04021},
  volume       = {33},
  year         = {2017},
}