Microfluidic antibody affinity profiling for in-solution characterisation of alloantibody – HLA interactions in human serum

Published on November 3rd, 2020

Authors: Matthias M. Schneider, Tom Scheidt, Ashley J. Priddey, Catherine K. Xu, Mengsha Hu, Sean R. A. Devenish, Georg Meisl, Christopher M. Dobson, Vasilis Kosmoliaptsis, and Tuomas P. J. Knowles

bioRxiv, 2020, 296442. DOI: 10.1101/2020.09.14.296442


The detection and characterization of antibodies in human blood is a key for clinical diagnostics and risk assessment for autoimmunity, infectious diseases and transplantation. Antibody titre derived from immunoassays is a commonly used measure for anti-body response, but this metric does not resolve readily the two fundamental properties of antibodies in solution, namely their affinity and concentration. This difficulty originates from the fact that the fundamental parameters describing the binding interaction, affinity and ligand concentration, are convoluted into the titre measurement; moreover, the difficulty of controlling the surface concentration and activity of the immobilized ligand can make it challenging to distinguish between avidity and affinity.

To address these challenges, Schneider et al. developed microfluidic antibody affinity profiling, an assay which allows the simultaneous determination of both affinity and antibody concentration, directly in solution, without surface immobilization or antibody purification. They demonstrate these measurements in the context of alloantibody characterization in organ transplantation, using complex patient sera, and quantify the concentration and affinity of alloantibodies against donor Human Leukocyte Antigens (HLA), an extensively used clinical biomarker to access the risk of allograft rejection. These results outline a path towards detection and in depth profiling of antibody response in patient sera.

Instrument: Fluidity One-W Serum
Therapeutic area: transplant

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