A Quantitative Holographic Agglutination Assay for Immunoglobulin A

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Hauptverfasser: Quddus, Rushna, Kirshenbaum, Kent, Grier, David G.
Format: Preprint
Veröffentlicht: 2025
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author Quddus, Rushna
Kirshenbaum, Kent
Grier, David G.
author_facet Quddus, Rushna
Kirshenbaum, Kent
Grier, David G.
contents This study introduces a Holographic Agglutination Assay for quantifying levels of the immunoglobulin protein IgA in biological samples. This is the first example of a label-free and bead-free assay that quantifies protein agglutinates by direct detection using Total Holographic Characterization. A proof-of-concept assay for human serum immunoglobulins is demonstrated using Jacalin, the galactose-specific plant lectin, to induce selective agglutination. By analyzing the size, refractive index, and number of particles in an assay sample, we obtain a reproducible and quantitative measurement of galactosylated immunoglobulins in a given sample. The assay is calibrated for a physiologically relevant reference interval of IgA concentrations in a 10x diluted emulated biological sample from low (80 mg/dL, 5 μM) to high (320 mg/dL, 20 μM) levels. The assay clearly distinguishes samples containing IgA from samples containing IgG. More broadly, this study introduces a platform for creating lectin-mediated Holographic Agglutination Assays to monitor levels of immunoglobulins in biological samples. The ability to quantify immunoglobulin levels efficiently in clinical samples is likely to be valuable for diagnostics and will provide a basis for assaying other proteins that can be induced to agglutinate.
format Preprint
id arxiv_https___arxiv_org_abs_2510_13480
institution arXiv
publishDate 2025
record_format arxiv
spellingShingle A Quantitative Holographic Agglutination Assay for Immunoglobulin A
Quddus, Rushna
Kirshenbaum, Kent
Grier, David G.
Optics
Medical Physics
Quantitative Methods
This study introduces a Holographic Agglutination Assay for quantifying levels of the immunoglobulin protein IgA in biological samples. This is the first example of a label-free and bead-free assay that quantifies protein agglutinates by direct detection using Total Holographic Characterization. A proof-of-concept assay for human serum immunoglobulins is demonstrated using Jacalin, the galactose-specific plant lectin, to induce selective agglutination. By analyzing the size, refractive index, and number of particles in an assay sample, we obtain a reproducible and quantitative measurement of galactosylated immunoglobulins in a given sample. The assay is calibrated for a physiologically relevant reference interval of IgA concentrations in a 10x diluted emulated biological sample from low (80 mg/dL, 5 μM) to high (320 mg/dL, 20 μM) levels. The assay clearly distinguishes samples containing IgA from samples containing IgG. More broadly, this study introduces a platform for creating lectin-mediated Holographic Agglutination Assays to monitor levels of immunoglobulins in biological samples. The ability to quantify immunoglobulin levels efficiently in clinical samples is likely to be valuable for diagnostics and will provide a basis for assaying other proteins that can be induced to agglutinate.
title A Quantitative Holographic Agglutination Assay for Immunoglobulin A
topic Optics
Medical Physics
Quantitative Methods
url https://arxiv.org/abs/2510.13480