Creative Biolabs is professional in biomarker assay development, validation, and profiling with multiplex biomarker panels, using immunoanalytical and enzymatic platforms, or LC-MS/MS, when appropriate. We have substantial experience with custom multiplex designs and can expertly help you design a custom biomarker assay to suit your needs.
Promising Multiplex Immunoassays
The simultaneous measurement of different targets from a single sample is an emerging area for achieving efficient and high-throughput detection. Multiplex immunoassays confer obvious advantages over widely used singleplex assays such as greater output per sample volume ratios, increased efficiency at a reduced expense, and higher throughput predicting more resolute. Contemporary multiplex immunoassay systems may be divided into two formats: planar assays and microbead-based suspension assays.
Fig.1 Multiplex formats in common use include planar-based assays or suspension-based assays.1
- For planar format, high-affinity capture ligands are immobilized discretely on a solid phase. The immobilized ligands are then exposed to treatment with the sample and probed with detection antibodies labeled with a reporter system.
- For suspension format, high-affinity capture ligands are immobilized discretely on fluorescently activated plastic microbeads and mixed with the sample in the liquid phase. Subsequent addition of detection antibodies labeled with a reporter dye enables high-resolution analysis of specific fluorescent signals via flow cytometric methods.
On-Demand Assay Design and Development
Our scientists will help to design and qualify study-specific biomarker assays. Although the ELISA is regarded as a singleplex assay format, the method can also be made suitable for multiplex protein analysis. Monoclonal, polyclonal, or recombinant antibodies with a defined specificity, sensitivity, and stability can be combined in one test to allow the binding of multiple proteins. We can transfer and qualify assays or develop them for you. Fast and accurate testing services are available once the assay is developed.
As a nimble biomarker CRO, we offer singleplex or multiplex assays on innovative platforms or as commercially available ELISA assays evaluated using a traditional 96-well plate format. All these assays undergo a fit-for-purpose biomarker validation, including assessing bioanalytical parameters such as accuracy, precision, and sensitivity. Once the biomarker assay is well characterized and robust, it can be relied upon to analyze preclinical samples. Specialized in biomarker assay development, we maintain high-quality standards in each step. Some of our most popular biomarker assay services include assay development, method validation, and cytokine testing.
Our biomarker platform offers several complementary solutions to fit every need you may have in terms of protein quantitation and profiling, as well as phosphorylation or glycosylation profiling, and mRNA/miRNA profiling. If you are interested in our multiple biomarker analysis services, please feel free to contact us for more information.
Published Data
1. Multiplex Immunoassay Platform for the Simultaneous Detection of Serum IgG Antibodies Targeting Six Human Coronaviruses
Fig.2 Correlation between monoplex and multiplex assay.2
In this study, a novel multiplexed magnetic microsphere immunoassay (MMIA) platform was developed and evaluated, enabling simultaneous IgG detection against six human coronaviruses (hCoVs) recombinant nucleocapsid proteins. Researchers employed paired human serum samples to assess IgG reactivity against six hCoVs, evaluating the sensitivity, specificity, and reproducibility of the assay. The results showed no signal interference occurred between monoplex and multiplex formats, with R² values ranging from 0.87 to 0.97. MMIA detected IgG antibodies with 86% sensitivity (92 of 106 positive samples) and 84% specificity (68 of 80 negative samples). This study demonstrates the feasibility of using MMIA as a platform for conducting large-scale seroprevalence studies of hCoVs.
2. Development of Multiplex Microsphere Immunoassays for Arboviral IgM and IgG Detection
Fig.3 IgM based on median fluorescent intensity (MFI) of sample reacted on viral antigen/negative antigen.3
This study developed a 13-virus multiplexed IgM and IgG microsphere immunoassay (MIA) for arboviruses, incorporating both internal and external controls. Traditional serodiagnosis for arboviruses often involves a combination of individual ELISAs and plaque-reduction neutralization tests, which can be cumbersome, especially when testing multiple viruses from specific geographic regions. The new multiplex assay allowed concurrent testing for multiple viruses, improving efficiency. The study evaluated 8 statistical classification methods to determine the best approach for analyzing results, with a focus on geographic regions. The "Logitboost" logistic regression method was identified as the most efficient for classification. When combining data from all tested samples, the error rates for the multiplex IgM and IgG MIAs were less than 5% for all geographic batteries. This work represents the most comprehensive, validated multiplexing method for arboviruses and offers a systematic approach to selecting optimal classification strategies for serologic testing.
References
- Ahsan, Haseeb, and Rizwan Ahmad. "Multiplex technology for biomarker immunoassays." Innate Immunity in Health and Disease (2020): 1-10.
- Trivedi, Suvang U., et al. "Development and evaluation of a multiplexed immunoassay for simultaneous detection of serum IgG antibodies to six human coronaviruses." Scientific reports 9.1 (2019): 1390. Distributed under Open Access license CC BY 4.0, without modification.
- Basile, Alison J., et al. "Multiplex microsphere immunoassays for the detection of IgM and IgG to arboviral diseases." PloS one 8.9 (2013): e75670. Distributed under Open Access license CC0 1.0.
For Research Use Only.
