Eleonora Macchia team at Åborough Academic University, Finland. Sensor: Optimized immunoglobulin detection and experimental technology design based on ultrasensitive sandwich immunoassay

In recent years, the identification of proteins, peptides, and genetic markers using femmole detection limits has become a fundamental task
for early diagnosis.
The detection of clinically relevant biomarkers at very low concentrations enables a better understanding of the etiology of many disease states and early diagnosis
of progressive and infectious diseases.
Standard medical analysis usually uses nucleic acids, proteins, or peptides as biomarkers
.
For genetic biomarkers, applications for clinical purposes (to distinguish between two different forms of leukemia) were carried out as early as 1999, and since then techniques
have been developed to identify oncogene alterations by different genomic markers.
Next-generation sequencing (NGS) technology has the most sensitive method for genetic markers with a limit of detection (LOD) for a single DNA or RNA copy, and has made significant progress
in the early detection of genetic markers for progressive diseases.
On the other hand, peptide and protein biomarkers also play a key role
in the early diagnosis of many pathological conditions.
Therefore, researchers have made great efforts in ultra-sensitive protein detection, but ultra-sensitive protein detection is still an extremely challenging task
.

Recently, Eleonora Macchia's team from Åbo Academic University in Finland published a research article in Advanced Sensor Research, reporting on the experimental
design of immunoglobulin detection to reduce the consumption of reagents and improve detection sensitivity.

In this work, an optimized Simoa planar array assay for the detection and quantification of immunoglobulin M (IgM, a nonspecific indicator of inflammation) was developed and optimized using a full-factor experimental design
.
Compared with the traditional single variable control method, the experimental design method in this paper reduces the overall experimental workload and improves the confidence of the experimental conclusions
.
By optimizing the capture antibody and detecting the concentration of the antibody, and through mathematical modeling and statistical analysis, the optimized parameters can achieve the detection limit of IgM up to 4 f_M
.
Compared to the commercially available Simoa Planar Array Kit, the protocol performance in this work is comparable and the cost of performing immunoglobulin assays is greatly reduced
as the required concentrations of the capture and detection layers in solution decrease by an order of magnitude.

According to the authors, this work uses a concise mathematical model to describe the factor-effect relationship of the detection process, and reduces the number of
experiments required through an experimental matrix and a reasonable hypothetical model.
The experimental results show that at lower detection antibody concentrations, changes in the concentration of captured antibodies do not significantly affect the detection limit
.
Therefore, the optimal conditions in the experimental domain correspond to the lowest acceptable detectable antibody concentration, combined with a reduction in the concentration of captured antibody to minimize assay cost
.

WILEY

Paper Information:

Implementation of Experimental Design Techniques to Optimize Immunoglobulins Detection with Ultrasensitive Sandwich Immunoassays

Cecilia Scandurra, Paolo Bollella, Ronald Österbacka, Francesco Leonetti, Eleonora Macchia, Luisa Torsi

Advanced Sensor Research

10.
1002/adsr.
202200009

Translated by Yichen Pan, Procter & Gamble

Click "Read Original" in the lower left corner to view the original text
of the paper.

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