Skip to main content

LUP Student Papers

LUND UNIVERSITY LIBRARIES

Development of a thiol-ene microfluidic Concanavalin A (ConA) functionalized chip for retention of glycoproteins and subsequent CE-UV analysis

Fürnsinn, Monika Rosa LU (2025) KBTM01 20251
Biotechnology (MSc)
Biotechnology (M.Sc.Eng.)
Abstract
The selective separation of glycoproteins is essential in biomarker discovery, glycoprotein profiling and analysis. Glycosylation of proteins has a major impact on their structure and function. The heterogeneity of glycosylation patterns makes the glycomic profiling extensive, but nonetheless of upmost importance for the deeper understanding of metabolic pathways, cell signalling and the impact of glycosylation alterations. The conventional methods for (glyco)protein analysis from biological samples, come with tedious sample preparation, that also have high reagent use and sometimes lead to loss of protein. By combining loading, washing and elution in one microfluidic device, loss of protein, sample preparation
time and reagent waste can... (More)
The selective separation of glycoproteins is essential in biomarker discovery, glycoprotein profiling and analysis. Glycosylation of proteins has a major impact on their structure and function. The heterogeneity of glycosylation patterns makes the glycomic profiling extensive, but nonetheless of upmost importance for the deeper understanding of metabolic pathways, cell signalling and the impact of glycosylation alterations. The conventional methods for (glyco)protein analysis from biological samples, come with tedious sample preparation, that also have high reagent use and sometimes lead to loss of protein. By combining loading, washing and elution in one microfluidic device, loss of protein, sample preparation
time and reagent waste can be minimized. This thesis work aimed to develop a microfluidic lectin affinity platform for the selective separation of glycoproteins, utilizing a Concanavalin A (ConA)-functionalized “monolithic” chip. The immobilisation of a non-enzymatic protein on a thiol-ene monolith chip is, to our knowledge, a novel approach. Using invertase as a model glycoprotein, the system is systematically characterized under varying sample concentrations, buffer and flow conditions to evaluate binding efficiency, elution performance and chip capacity. Despite the variability in yield between the chip replicates the system demonstrated the ability to bind invertase and recovering it via competitive elution with methyl-α-D mannopyranoside. Two different channel designs were tested, and their performance and flow characteristics were compared to each other. The long channel chip (LCC) with a smaller cross section resulted in higher recovery yield than the short channel chip (SCC), however pressure drop calculations
reveal that the operating flow rates for the LCC have to be around 2 µL/min to not disrupt the monolith structure in the channel. The maximum amount of invertase eluted off the LCC and SCC was measured with CE-UV to be ~20 pmol and ~15 pmol, respectively.
Overall, this work presents a promising step toward integrating lectin affinity chromatography into microfluidic systems for glycoprotein handling. The immobilization of ConA on a thiol-ene monolith and the demonstrated binding of a model glycoprotein mark an advance toward miniaturized, efficient sample preparation workflows. While further optimization of the system is required to enhance reproducibility and binding capacity, the platform lays the groundwork for future applications. (Less)
Popular Abstract
Proteins often carry sugar molecules that play an essential role in how cells communicate, respond to their environment, and function properly. These “sugar-coated” proteins are called glycoproteins. The patterns of the sugar chains on the protein surface are unique. Changes in these sugar patterns can be early warning signs for many diseases, including cancer, making them a valuable target for medical research. However, studying glycoproteins is complex: traditional methods for separating and analyzing glycoproteins are slow, wasteful, and often loose some of the proteins along the way.
This thesis explores a new small microfluidic chip device that can capture glycoproteins using a sugar binding protein called Concanavalin A (ConA).... (More)
Proteins often carry sugar molecules that play an essential role in how cells communicate, respond to their environment, and function properly. These “sugar-coated” proteins are called glycoproteins. The patterns of the sugar chains on the protein surface are unique. Changes in these sugar patterns can be early warning signs for many diseases, including cancer, making them a valuable target for medical research. However, studying glycoproteins is complex: traditional methods for separating and analyzing glycoproteins are slow, wasteful, and often loose some of the proteins along the way.
This thesis explores a new small microfluidic chip device that can capture glycoproteins using a sugar binding protein called Concanavalin A (ConA). Inside the chip channel there is a sponge-like material (a monolith), which increases the surface area for capturing proteins. On this monolith the ConA is permanently fixed and binds the glycoproteins while they pass by. This small device helps capture the
glycoproteins efficiently, using less time, fewer chemicals and smaller samples than traditional methods.
Employing invertase, a glycoprotein, as a model protein, the system’s performance was tested under different conditions. For example, different designs of the chip channel were examined to see which worked best. The design featuring a longer channel was better at collecting the invertase but required very slow flow to avoid damaging the monolith inside the channel.
In conclusion, this work is a step forward towards fixing a lectin inside the monolithic channel in order to isolate glycoproteins on a small scale. While more development is needed to improve reliability, the device offers a foundation for future tools in biomedical research and diagnostics. (Less)
Please use this url to cite or link to this publication:
author
Fürnsinn, Monika Rosa LU
supervisor
organization
course
KBTM01 20251
year
type
H2 - Master's Degree (Two Years)
subject
keywords
Lectin-functionalized microfluidic device, Thiol-ene monolith, Concanavalin A, Invertase, Methyl-α-D-mannopyranoside, Capillary electrophoresis, Design and flow optimisation, Biotechnology
language
English
id
9206099
date added to LUP
2025-06-27 11:42:34
date last changed
2025-06-27 11:42:34
@misc{9206099,
  abstract     = {{The selective separation of glycoproteins is essential in biomarker discovery, glycoprotein profiling and analysis. Glycosylation of proteins has a major impact on their structure and function. The heterogeneity of glycosylation patterns makes the glycomic profiling extensive, but nonetheless of upmost importance for the deeper understanding of metabolic pathways, cell signalling and the impact of glycosylation alterations. The conventional methods for (glyco)protein analysis from biological samples, come with tedious sample preparation, that also have high reagent use and sometimes lead to loss of protein. By combining loading, washing and elution in one microfluidic device, loss of protein, sample preparation 
time and reagent waste can be minimized. This thesis work aimed to develop a microfluidic lectin affinity platform for the selective separation of glycoproteins, utilizing a Concanavalin A (ConA)-functionalized “monolithic” chip. The immobilisation of a non-enzymatic protein on a thiol-ene monolith chip is, to our knowledge, a novel approach. Using invertase as a model glycoprotein, the system is systematically characterized under varying sample concentrations, buffer and flow conditions to evaluate binding efficiency, elution performance and chip capacity. Despite the variability in yield between the chip replicates the system demonstrated the ability to bind invertase and recovering it via competitive elution with methyl-α-D mannopyranoside. Two different channel designs were tested, and their performance and flow characteristics were compared to each other. The long channel chip (LCC) with a smaller cross section resulted in higher recovery yield than the short channel chip (SCC), however pressure drop calculations 
reveal that the operating flow rates for the LCC have to be around 2 µL/min to not disrupt the monolith structure in the channel. The maximum amount of invertase eluted off the LCC and SCC was measured with CE-UV to be ~20 pmol and ~15 pmol, respectively. 
Overall, this work presents a promising step toward integrating lectin affinity chromatography into microfluidic systems for glycoprotein handling. The immobilization of ConA on a thiol-ene monolith and the demonstrated binding of a model glycoprotein mark an advance toward miniaturized, efficient sample preparation workflows. While further optimization of the system is required to enhance reproducibility and binding capacity, the platform lays the groundwork for future applications.}},
  author       = {{Fürnsinn, Monika Rosa}},
  language     = {{eng}},
  note         = {{Student Paper}},
  title        = {{Development of a thiol-ene microfluidic Concanavalin A (ConA) functionalized chip for retention of glycoproteins and subsequent CE-UV analysis}},
  year         = {{2025}},
}