Absolute Quantitation of Serum Antibody Reactivity Using the Richards Growth Model for Antigen Microspot Titration.
Krisztian PappÁgnes KovácsAnita OroszZoltán HérincsJudit RandekKároly LiliomTamás PfeilJózsef PrechlPublished in: Sensors (Basel, Switzerland) (2022)
In spite of its pivotal role in the characterization of humoral immunity, there is no accepted method for the absolute quantitation of antigen-specific serum antibodies. We devised a novel method to quantify polyclonal antibody reactivity, which exploits protein microspot assays and employs a novel analytical approach. Microarrays with a density series of disease-specific antigens were treated with different serum dilutions and developed for IgG and IgA binding. By fitting the binding data of both dilution series to a product of two generalized logistic functions, we obtained estimates of antibody reactivity of two immunoglobulin classes simultaneously. These estimates are the antigen concentrations required for reaching the inflection point of thermodynamic activity coefficient of antibodies and the limiting activity coefficient of antigen. By providing universal chemical units, this approach may improve the standardization of serological testing, the quality control of antibodies and the quantitative mapping of the antibody-antigen interaction space.
Keyphrases
- quality control
- liquid chromatography
- liquid chromatography tandem mass spectrometry
- mass spectrometry
- high resolution
- immune response
- electronic health record
- high performance liquid chromatography
- high throughput
- tandem mass spectrometry
- magnetic resonance imaging
- diffusion weighted imaging
- dna binding
- machine learning
- artificial intelligence
- big data
- protein protein
- transcription factor