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  Direct Detection of Antibody Concentration and Affinity in Human Serum Using Microscale Thermophoresis

Lippok, S., Seidel, S. A. I., Duhr, S., Uhland, K., Holthoff, H.-P., Jenne, D. E., et al. (2012). Direct Detection of Antibody Concentration and Affinity in Human Serum Using Microscale Thermophoresis. ANALYTICAL CHEMISTRY, 84(8), 3523-3530. doi:10.1021/ac202923j.

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 Creators:
Lippok, Svenja1, Author
Seidel, Susanne A. I.1, Author
Duhr, Stefan1, Author
Uhland, Kerstin1, Author
Holthoff, Hans-Peter1, Author
Jenne, Dieter E.2, Author           
Braun, Dieter1, Author
Affiliations:
1External Organizations, ou_persistent22              
2Department: Neuroimmunology / Wekerle, MPI of Neurobiology, Max Planck Society, ou_1113547              

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Free keywords: IDIOPATHIC DILATED CARDIOMYOPATHY; FORCED RAYLEIGH-SCATTERING; BETA-ADRENERGIC-RECEPTOR; HEART-FAILURE; BETA(1)-ADRENERGIC RECEPTORS; AUTOANTIBODIES; BINDING; ASSAYS
 Abstract: The direct quantification of both the binding affinity and absolute concentration of disease-related biomarkers in biological fluids is particularly beneficial for differential diagnosis and therapy monitoring. Here, we extend microscale thermophoresis to target immunological questions. Optically generated thermal gradients were used to deplete fluorescently marked antigens in 2- and 10-fold-diluted human serum. We devised and validated an autocompetitive strategy to independently fit the concentration and dissociation constant of autoimmune antibodies against the cardiac beta 1-adrenergic receptor related to dilated cardiomyopathy. As an artificial antigen, the peptide CORI was designed to mimic the second extracellular receptor loop. Thermophoresis resolved antibody concentrations from 2 to 200 nM and measured the dissociation constant as 75 nM. The approach quantifies antibody binding in its native serum environment within microliter volumes and without any surface attachments. The simplicity of the mix and probe protocol minimizes systematic errors, making thermophoresis a promising detection method for personalized medicine.

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Language(s): eng - English
 Dates: 2012-04-17
 Publication Status: Issued
 Pages: 8
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Identifiers: ISI: 000302838500007
DOI: 10.1021/ac202923j
 Degree: -

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Title: ANALYTICAL CHEMISTRY
Source Genre: Journal
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Publ. Info: 1155 16TH ST, NW, WASHINGTON, DC 20036 USA : AMER CHEMICAL SOC
Pages: - Volume / Issue: 84 (8) Sequence Number: - Start / End Page: 3523 - 3530 Identifier: ISSN: 0003-2700