How Sweet Can Multiplexing Be? Profiling Cancer-Related Autoantibodies using Glycan-Coated Microspheres

Presented at: xMAP Connect
Speakers

Abstract:

Background and aim:
Glycosylation is an essential posttranslational modification used primarily for cellular communication. The formation of complex glycans as secondary gene products is dependent on the activity of various glycosyltransferases (GTs). Mutated GTs, however, can cause significant alterations in the glycan patterns expressed on cancerous cells, making those patterns potent biomarkers. Whereas the detection of these tumor-associated carbohydrate antigens (TACA) require primary tumor tissue, autoantibodies (AAbs) against cancer-specific glycans in human plasma can be detected with a simple and minimal invasive technique.

Methods:
The Glyco-Multiplex assay enables the simultaneous detection of AAbs against (i) various glycans with (ii) minor amounts of plasma from (iii) a large number of patients in (iv) a short time. For this approach, multiple, potentially tumor-associated, glycans are coupled to distinct Luminex® bead regions. The unique color-coding of the beads allows the measurement of an individual’s isotypespecific AAb profile against the selected glycans simultaneously in one single well. The adaption of the coupling protocol for the use of the KingFisherTM Flex enables the upscale of this assay to up to 96 different regions with negligible additional hands-on time.

Results:
A protocol for the coupling of glycans to carboxylated MagPlex® beads was established and optimized. Subsequently, suitable combinations of plasma concentrations and Ig-specific (IgM, IgG and IgA) detection antibodies and assay controls were evaluated to detect α-TACA AAbs in human plasma samples. To get a first impression on α-TACA AAb signals, plasma samples from colon cancer patients, lung cancer patients and healthy donors were measured using the Glyco-Multiplex. In accordance with literature, (auto)antibodies in both healthy and diseased individuals could be detected. Furthermore, the analysis revealed individual isotype (IgM, IgG, IgA)-specific α-TACA-AAbs profiles for the tested samples. MagPlex® beads that were coupled using the protocol for the KingFisherTM Flex yielded results that were comparable to those obtained with manually coupled beads.

Conclusion:
Taken together, we are now able to generate individual autoantibody profiles against glycan structures, using minor amounts of plasma, in a high throughput manner.


Show Resources
You May Also Like
SEP 05, 2019 04:00 PM CEST
C.E. CREDITS
SEP 05, 2019 04:00 PM CEST
DATE: September 5, 2019TIME: 7:00am PT, 10:00am ET, 4:00pm CEST PCR (Polymerase Chain Reaction) has gone through a massive evolution since its development in 1983. Besides it...
AUG 27, 2019 09:00 AM PDT
C.E. CREDITS
AUG 27, 2019 09:00 AM PDT
DATE: August 27, 2019 TIME: 9:00am PDT, 12:00pm EDT Immunotherapies targeting PD-1 or PD-L1 have proven remarkably effective for treating cancer in some patients, with considerabl...
JAN 23, 2020 09:00 AM PST
Add to Calendar Select one of the following: iCal Google Calendar Outlook Calendar Yahoo Calendar
C.E. CREDITS
JAN 23, 2020 09:00 AM PST
Add to Calendar Select one of the following: iCal Google Calendar Outlook Calendar Yahoo Calendar
DATE: January 23, 2020 TIME: 9:00am PST, 12:00pm EST...
JUL 31, 2019 09:00 AM PDT
C.E. CREDITS
JUL 31, 2019 09:00 AM PDT
DATE: July 31, 2019TIME: 9:00am PT, 12:00pm ET The choroid plexus, which makes up the blood-cerebrospinal fluid barrier in the central nervous system (CNS), lines the ventricle...
AUG 15, 2019 09:00 AM PDT
C.E. CREDITS
AUG 15, 2019 09:00 AM PDT
DATE: August 15, 2019TIME: 9:00am PDT, 12:00pm EDT Radiation therapy is a critical tool for the treatment of brain tumors, however, exposure to high doses of ionizing radiation...
AUG 13, 2019 09:00 AM PDT
C.E. CREDITS
AUG 13, 2019 09:00 AM PDT
DATE: August 13, 2019TIME: 9:00am PT, 12:00pm ET, 5:00pm BST Molecular complexes are major constituents of cells, hence unraveling their mechanisms is key to fuller comprehension of c...
Loading Comments...
Show Resources
Event Countdown
  • 0 Days
  • 0 Hours
  • 0 Minutes
  • 0 Seconds