xMAP Multiplexing – Page 11 – Diasorin & Luminex Support

MAGPIX^® Systems See New Life Through Equipment Donation Program

MilliporeSigma and Seeding Labs send gently used instruments to labs in need

Thanks to a nonprofit organization called Seeding Labs and our Licensed Technologies Partner MilliporeSigma, previously-owned ® Systems” href=”https://www.luminexcorp.com/research-magpix/#overview”>MAGPIX^® Systems have gotten new life through this program; Vaughn has already facilitated the donation of several of them, which found a second home at university labs in Africa, and has more in the donation pipeline. In addition to helping scientists build research infrastructure in underfunded labs, this program also helps researchers who want to upgrade their equipment without sending older systems to a landfill.

MAGPIX^® Systems are a good fit for the Seeding Labs program, according to Vaughn, because they’re simple instruments that require minimal maintenance and can be easily installed by users. Because Luminex and MilliporeSigma offer so many video tutorials for working with these systems, like on Seeding Labs’ TeleScience platform, researchers anywhere can easily get them up and running, troubleshoot basic problems, and learn to build their own multiplex assays. “It helps them get great results the first time out,” Vaughn says.

Resources

Luminex Licensed Technologies Partner Spotlight: MilliporeSigma [Blog]
Webinar Wednesday: For Tracking Anti-Apoptosis Proteins, Scientists Turn to Multiplexing [Blog]
Luminex MAGPIX System [Video]
Luminex xMAP Technology Overview [Video]
Getting Started with xMAP Technology [Video]

METHODS Journal: A New Multiplexing Protocol Performs Protein and Gene Level Measurements on a Single Luminex Platform

Researchers demonstrate a high-throughput method for measuring mRNA and proteins using the Luminex xMAP^® Platform

You shouldn’t have to choose between analyzing gene expression or screening for proteins. In a Methods publication, researchers demonstrate how to measure mRNA expression and analyze proteins with a single sample set using Luminex Advances in Bead-Based Biomarker Detection [Methods xMAP Issue]

® Cookbook to Design Your Own Assays” href=”http://info.luminexcorp.com/en-us/research/download-the-xmap-cookbook”>xMAP^® Cookbook to Design Your Own Assays [Download]

Webinar: Fast, Economical Malaria Testing with xMAP^® Technology

Multiplex assay allows for detailed investigations of transmission and drug resistance

Multiplexing technology from Luminex has been critical for the development of assays designed to study human SNPs involved in parasite transmission as well as the spread of drug resistance markers in Plasmodium falciparum. That was a key message in this presentation from Lynn Grignard, a research fellow at the London School of Hygiene and Tropical Medicine.

Malaria, which led to 212 million cases worldwide in 2015, is caused by a parasite with a remarkably complex life cycle. Important steps include development in the mosquito and transmission to a human host when the mosquito feeds, followed by a liver cycle and blood stage within the human. While malaria is preventable and curable, Grignard said, it remains a life-threatening disease, particularly in Africa.

Developing Assays

In her presentation, Grignard focused on two areas she studies: how human SNPs are involved in transmission or protection against malaria, and how drug resistance is spread to the parasite. These investigations led to the development of two assays based on ® Technology” href=”https://www.youtube.com/watch?v=m6DvXhtFOm4″>Getting Started with xMAP^® Technology [Video]

® Cookbook to Design Your Own Assays” href=”http://info.luminexcorp.com/en-us/research/download-the-xmap-cookbook”>xMAP^® Cookbook to Design Your Own Assays [Download]

An Expert’s Guide to Multiplexing Immunoassays [Blog]

Multiplex Malaria MagPlex-TAG Assay to Detect Human Host and Parasite Genetic Markers

Webinar: Multiplex Serological Assay Developed to Identify Avian Influenza Subtypes

Rapid detection could help prevent transmission and reduce flock losses

In addition to being a public health threat, avian influenza is also costly and potentially devastating to people whose livelihoods depend on raising poultry. To enable faster detection of bird flu and identification of its subtypes, scientists at Wageningen Bioveterinary Research in The Netherlands developed a multiplex assay using this 20-minute video presentation, veterinarian Evelien Germeraad reports on the development and validation of that assay. Wageningen Bioveterinary Research, part of Wageningen University & Research, is a national reference lab for infectious animal diseases, receiving samples for testing from all over the country. The lab tests cattle, poultry, goats, and other types of animals for illnesses ranging from rabies and tularemia to psittacosis and foot-and-mouth disease. Scientists there run as many as 300,000 assays per year, of which 20,000 are deployed for avian influenza, the main focus of Germeraad’s talk.

Poultry Industry in The Netherlands

In The Netherlands, the poultry industry is valued at about €1.5 billion. The country is home to 100 million chickens, and bird flu is not uncommon. Recent outbreaks occurred in 2003, 2014, and 2016. In the 2014 outbreak, which affected just a handful of farms, nearly 250,000 chickens had to be culled to contain the pathogen. This enormous scale of loss is why it’s so important to diagnose avian influenza quickly, Germeraad says.

One of the challenges of testing for avian influenza is the broad range of subtypes. There are 16 types of hemagglutinin in birds, plus nine types of neuraminidase, making for a large number of H+N protein subtypes of the virus. Some strains have low pathogenicity, while others are so severe that mortality can rise to 100% in just a few days, Germeraad says. Conventional serological assays — such as ELISA, hemagglutination inhibition testing, and agar gel immunodiffusion — are time-consuming, require high volumes of sample, and do not all produce subtype-specific results.

For a faster test that would identify all subtypes, Germeraad and her team turned to xMAP Technology and the ® Technology” href=”https://www.youtube.com/watch?v=m6DvXhtFOm4″>Getting Started with xMAP^® Technology [Video]

® Cookbook to Design Your Own Assays” href=”http://info.luminexcorp.com/en-us/research/download-the-xmap-cookbook”>xMAP^® Cookbook to Design Your Own Assays [Download]

An Expert’s Guide to Multiplexing Immunoassays [Blog]

Development of A Multiplex Serological Assay for Avian Influenza [VIEW NOW]

METHODS Special Issue: Multiplex Assays Enable Simultaneous Measurement of RNA and Proteins

A more comprehensive view sheds light on the biological response to muscle injury

Scientists at the University of North Texas have established a novel method for measuring a broad range of RNA and protein biomarkers. In their study, they employed bead-based multiplex detection to assess the effects of skeletal muscle injury over time, but the same concept could be useful for many clinical applications.

The team—led by Melody Gary—published their results in an April 2019 issue of Methods.

Multiplex methods can do more with less

For this project, they used a multiplex assay based on Advances in Bead-Based Biomarker Detection [Methods xMAP^® Issue]

® Technology” href=”https://www.youtube.com/watch?v=m6DvXhtFOm4″>Getting Started with xMAP^® Technology [Video]

® Kit Finder” href=”https://www.luminexcorp.com/kitfinder/”>Browse 1,200+ Partner Kits with xMAP^® Kit Finder [Online Tool]

METHODS Special Issue: Multiplex Assays Enable Simultaneous Measurement of RNA and Proteins

METHODS Special Issue: Multiplexing Protocol for Evaluating Biological Interactions with Nanoparticles

A new multiplexing protocol offers guidelines for measuring immunomodulatory effects of nanoparticles used in clinical applications

Nanoparticles have the potential to deliver more targeted therapies to the exact location in the body where they’re most needed. To do this, it is essential to understand how these foreign particles interact with a patient’s body. A recent publication in the Methods journal is a big step forward, offering a protocol for evaluating the body’s immune response to nanotechnology.

Scientists from Texas A&M University and Bio-Rad Laboratories designed parameters for evaluating the immunomodulatory effects of nano- and microparticles using multiplex biomarker analysis. To detect biomarkers of interest, the researchers utilized Advances in Bead-Based Biomarker Detection [Methods xMAP^® Issue]

Immunogenicity Profiling of Nanocarriers [xMAP^® Connect Webinar]

® Cookbook to Design Your Own Assays” href=”http://info.luminexcorp.com/en-us/research/download-the-xmap-cookbook”>xMAP^® Cookbook to Design Your Own Assays [Download]

METHODS Special Issue: Multiplexing Protocol for Evaluating Biological Interactions with Nanoparticles

METHODS Special Issue: Multiplexing for Personalized Oncology

Assays quantify RNA expression even in challenging samples

Multiplex molecular assays have several important roles in the realm of personalized cancer medicine. A team of researchers at the University of Malta recently published a paper, titled “Bead-based RNA multiplex panels for biomarker detection in oncology samples,” in the Methods journal that offers an overview of those roles, showing that multiplex assays based on QuantiGene™ Plex Assay from Thermo Fisher which allows customization to quantify expression of as many as 80 genes, the scientists report that large validation studies have established the accuracy and precision of a multiplex approach. Compared to RT-PCR, the multiplex assay had better sensitivity, linearity, precision, and relative accuracy. It was also more sensitive for genes with low expression compared to a number of array-based methods.

The authors note that multiplex assays work well with stained and unstained tissue sections, so they can even be used after traditional histological analysis has been conducted. Further, the assays have been successful for liquid biopsy analyses, detecting circulating tumor cells that can provide valuable insight for patient monitoring. Thanks to the large number of genes that can be assayed at once, this method can help characterize the heterogeneity of a tumor as well.

“High sensitivity and specificity, together with multiplexing ability, make bead-based RNA assays highly useful for biomarker detection in oncology samples,” the scientists write. “We suggest that this method is a preferable option for multiplexing expression analysis of up to 80 genes, especially when using highly degraded specimens or low sample concentrations.”

Resources

Advances in Bead-Based Biomarker Detection [Methods xMAP Issue]
® Cookbook to Design Your Own Assays” href=”http://info.luminexcorp.com/en-us/research/download-the-xmap-cookbook”>xMAP^® Cookbook to Design Your Own Assays [Download]

Webinar: Improving Rheumatoid Arthritis Diagnostics with Multiplex Assays

How a European team avoided performing 16,000 ELISA tests

New efforts to improve the diagnosis of rheumatoid arthritis, as well as to guide treatment selection, are making strong progress thanks to presentation from Erik Lönnblom of the Karolinska Institute. Multiplex immunoassays have allowed his team and collaborators to generate 1.5 million data points so far — a feat that would otherwise have required nearly 16,000 ELISA plates to achieve.

Lönnblom works in a lab focused on characterizing epitopes to type II collagen, which is a candidate autoantigen for rheumatoid arthritis. The protein, which is quite difficult to work with, is highly conserved; epitopes are virtually identical across mice, rats, humans, and monkeys. One of the most promising areas of study is a post-translational modification known as citrullination. Anti-citrulline protein antibodies are present in more than two-thirds of patients with rheumatoid arthritis, and they can be detected years before the onset of clinical symptoms of the disease.

Making Strides with Rheumatoid Arthritis

There are several diagnostic challenges relevant to rheumatoid arthritis. Lönnblom says the most pressing areas include earlier and more accurate disease diagnosis; prediction of response to treatment; and prediction of disease onset in healthy individuals. As part of a multi-institution consortium, Lönnblom and his colleagues have already made inroads.

That would not have happened so quickly without xMAP Technology for multiplex assays. Originally, Lönnblom says, the plan was to use ELISAs for everything. But as the scientists calculated the number of assays they’d need to run to study the 400 proteins or peptides relevant to rheumatoid arthritis, it became clear the approach was not feasible. Sample requirements alone were a deal-breaker: while each Luminex assay needs just 1 microliter of material, Lönnblom says, ELISA tests for the same biomarkers would consume half a milliliter or more, an amount that no clinical sample repository could part with.

So far, the team has run the multiplex assay on several thousand clinical samples. Already, important insights are emerging. One promising new biomarker appears to predict some established cases of rheumatoid arthritis more reliably than previously known biomarkers. Another seems to predict poor response to a specific therapy, essential information for the grueling trial-and-error method of therapies used for patients today.

Continuing Research

Looking ahead, Lönnblom and his team plan to perform technical validation of the most promising peptides, as well as clinical validation of their current findings. They also aim to build more sophisticated diagnostic models for improved treatment of patients with rheumatoid arthritis.

Resources

® Kit Finder” href=”https://www.luminexcorp.com/kitfinder/”>Browse 1,200+ Partner Kits with xMAP^® Kit Finder [Online Tool]
An Expert’s Guide to Multiplexing Immunoassays [Blog]

METHODS Special Issue: Multiplexing to Quantify Cytokines and Chemokines in Mouse Brain Study

Assay requires minimal sample volume for 33-analyte interrogation

Scientists from the National Institute of Neurological Disorders and Stroke and Bio-Rad Laboratories published a helpful protocol for a method to analyze more than 30 cytokines and chemokines in mouse brain tissue. The technique can be adapted to other sample types as well.

The paper, titled “Method to quantify cytokines and chemokines in mouse brain tissue using ® Technology” href=”https://www.luminexcorp.com/xmap-technology/”>xMAP^® Technology.

“These assays allow researchers to analyze an entire network of cytokines/chemokines in a single sample, which conserves tissue, streamlines workflows, and accelerates research,” the scientists note. “In addition to saving time and money, multiplex immunoassays help advance vaccine development, drug discovery, basic research, and clinical trials by providing a quantitative snapshot of immune mediators expressed in samples of interest.”

For this project, the team focused on Plasmodium berghei, a parasite that causes cerebral malaria in mice. Two groups of mice—those with and without CD8+ T cells—were infected with the pathogen, and a third group was used as a control. Subsequently, they analyzed the expression of 33 chemokines and cytokines using the Bio-Plex^® multiplex immunoassay to characterize the inflammatory profile, as well as the role of CD8+ T cells.

“We observed elevated chemokine and cytokine levels in the brains of infected mice during development of cerebral malaria,” the scientists report. “Twenty different analytes were upregulated, with the majority being chemokines involved in the recruitment [of] immune cells.” They also found that CD8+ T cells are important for promoting inflammation at the onset of cerebral malaria.

The paper also includes a discussion about the utility and logistics of multiplex immunoassays. The approach was successful in overcoming the sample volume obstacle: scientists needed just 50 µl of tissue sample from each mouse to evaluate all 33 chemokines and cytokines in a single assay. Authors also encourage anyone planning to try this protocol to ensure detailed planning and proper timing of all steps. “For optimal results, it is important to plan and allow sufficient time to perform instrument validation/calibration, design plate layouts, and perform mixing/dispensing steps with precision,” they write.

Resources

Advances in Bead-Based Biomarker Detection [Methods xMAP Issue]
® Cookbook to Design Your Own Assays” href=”http://info.luminexcorp.com/en-us/research/download-the-xmap-cookbook”>xMAP^® Cookbook to Design Your Own Assays [Download]

METHODS Special Issue: New Multiplex Assay for Vaccine Immunogenicity Testing

Recently developed test lowers cost and reduces sample volumes needed for evaluating combination vaccines

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Scientists at the Serum Institute of India have developed a new multiplex immunoassay to perform the mouse immunogenicity testing recommended in many countries for quality control in the production of combination vaccines. They chose xMAP^® Technology to design the serological assay, which features higher throughput, faster analysis, and lower sample volume compared to serial ELISA testing.

How the Serological Assay Works

The team reported this work in a recent publication in Methods. The pentaplex assay they created “simultaneously measures the concentration of serum (IgG) antibodies against B. Pertussis antigens; pertussis toxin, filamentous hemagglutinin (FHA), pertactin (PRN) and tetanus (T), and diphtheria (D) toxoid in the Tdap vaccine-immunized animals,” the authors write.

While combination vaccines are important for a host of reasons, preclinical evaluation of them can be challenging. Even later in the development phase, these multivalent vaccines can be difficult to monitor. For that reason, the World Health Organization and European Pharmacopeia both encourage non-lethal immunogenicity testing in mice “for monitoring lot-to-lot consistency during manufacturing,” the scientists note. In conventional testing, “the assay involves immunization of mice with serial dilutions of the vaccine, [where the] serum IgG antibody response against each pertussis vaccine component is then measured by [ELISA].”

Multiplexing to Reduce Cost and Time

This multiplexing project was launched to eliminate the need for ELISAs, which are costly, tedious to run, and require a significant amount of serum. The team selected xMAP Technology due to its prior success in this area: “Several studies have previously described the accuracy and high-throughput advantage of this platform for evaluating vaccine-elicited binding antibodies,” the authors report.

After development, the multiplex assay was evaluated using serum samples taken from animals with varying dilutions of immunization. “The assay was able to predict the dose-dependent trend in IgG responses against all five antigens, which will be important to define the potency and quality of [each] vaccine against a reference vaccine batch of proven clinical efficacy,” the scientists note. Validation was performed according to international regulatory guidelines.

“The assay described here meets all the requirements of specificity, selectivity, reproducibility, and accuracy, and is thus a viable alternative to conventional ELISA for detection of DTaP3 antibodies in mouse serum samples,” the authors conclude.

Resources

Bead-Based Multiplexing for Immunogenicity Testing [Web page]
At the Serum Institute of India, Vaccine Development Requires Multiplex Assays [Blog]
Advances in Bead-Based Biomarker Detection [Methods xMAP Issue]
® Cookbook to Design Your Own Assays” href=”http://info.luminexcorp.com/en-us/research/download-the-xmap-cookbook”>xMAP^® Cookbook to Design Your Own Assays [Download]
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MilliporeSigma and Seeding Labs send gently used instruments to labs in need

Resources

Researchers demonstrate a high-throughput method for measuring mRNA and proteins using the Luminex xMAP® Platform

Multiplex assay allows for detailed investigations of transmission and drug resistance

Developing Assays

Rapid detection could help prevent transmission and reduce flock losses

Poultry Industry in The Netherlands

A more comprehensive view sheds light on the biological response to muscle injury

Multiplex methods can do more with less

A new multiplexing protocol offers guidelines for measuring immunomodulatory effects of nanoparticles used in clinical applications

Assays quantify RNA expression even in challenging samples

Resources

How a European team avoided performing 16,000 ELISA tests

Making Strides with Rheumatoid Arthritis

Continuing Research

Resources

Assay requires minimal sample volume for 33-analyte interrogation

Resources

Recently developed test lowers cost and reduces sample volumes needed for evaluating combination vaccines

How the Serological Assay Works

Multiplexing to Reduce Cost and Time

Resources

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Our Solutions

Useful Links

Legal Information

Document Repository

Researchers demonstrate a high-throughput method for measuring mRNA and proteins using the Luminex xMAP^® Platform