Overview

Neurodegenerative diseases (ND) affect millions of people worldwide and are characterized by the accumulation of neurotoxic amyloid aggregates in the brain leading to loss of function and ultimately death of neuronal cells. Immunotherapeutic approaches against NDs focus on antibodies that target the pathogenic aggregated forms of proteins such as α-synuclein or Aß peptide related to Parkinson’s disease and Alzheimer’s disease, respectively. ELISA and other surface-based methods are commonly used to identify antibodies that specifically target these pathogenic aggregates but tethering these sensitive proteins to a surface is often not feasible or induces artificial structures that do not occur in patients. Microfluidic diffusional sizing (MDS) avoids surface immobilization and thus preserves the disease-relevant structures of the aggregates which enables identification of antibodies that bind with high specificity and have the most effective mode of action against disease progression.

Purification-free quantification of affinity and receptor expression levels.

Fig 1. Overview of the purification-free workflow offered by the MAffCon Assay.

Case Study

Purification-free affinity and concentration measurement of membrane-protein targets

In this collaboration with the University of Graz, the MAffCon Assay was used to measure the binding of trastuzumab (Herceptin) to HER2 extracted from breast cancer cells, with consistent results to literature values and the potential for straightforward quantification of HER2 expression. 

“

The improvement using MDS comes from the fact that we can produce this robust and precise data without much purification.

Lena Bauernhofer

Institute of Molecular Biosciences, University of Graz

Read the application note

Get started

To study membrane proteins without purification, we recommend the following:

Membrane Proteins

Fluidity One-M

Fluidity Intelligence

Results

Affinity, Receptor Expression Levels, Concentration

Workflow specification and benefits:

• 2 hours of sample preparation + overnight incubation (less than 30 minutes hands-on time)  
• 2-3 hours for data collection and analysis 
• K_D range from pM to µM  
• Concentration ranges from nM to µM 
• Provides expression levels in terms of receptors per cell 
• One standard T175 cell culture flask at 90–95% confluency will be sufficient for several experiments 
• 12 µL of sample per triplicate  
• Quick & easy to perform