In our final blog post on double-stranded RNA (dsRNA), we turn our attention to the chemical building blocks of mRNA therapeuticsโmodified nucleotides. These seemingly minor changes to the RNA sequence play a crucial role in the success of mRNA-based vaccines and treatments. However, they also introduce complexities in accurately detecting and quantifying unwanted dsRNA byproductsโ key steps in ensuring the therapeutic efficacy of your mRNA product.
What Are Modified Nucleotides?
Modified nucleotides are ribonucleotides containing chemically altered nucleosides — like specialty ingredients swapped into a classic recipe to improve taste and nutrition. Just as a chef might use a lactose-free milk or gluten-free flour to make a dish easier to digest without changing its core structure, scientists use chemically altered nucleosides during in vitro transcription (IVT) to improve how mRNA therapies perform. These modifications replace their natural counterparts (e.g., uridine or cytidine) in the final RNA product. Their incorporation improves the performance and safety of mRNA therapeutics in several ways:
As mRNA therapeutics continue to expand across clinical pipelines, one persistent challenge remains for developers: reducing double-stranded RNA (dsRNA) contaminants that can compromise safety and efficacy. These unintended byproducts of in vitro transcription (IVT) can trigger unwanted immune responses and reduce the potency of the final product. Developers must prioritize dsRNA detection and control as essential steps in the process. In our previous blog post we offered a high-level discussion of what is double-stranded RNA (dsRNA), its biological function, and importance of detection in a therapeutic context. Here, weโll take a closer look at origins of dsRNA contamination, quality control measures, and improvement strategies.
Large-scale production of single-stranded RNA (ssRNA) for mRNA-based therapeutics is primarily done through in vitro transcription (IVT), an enzymatic process designed to generate high-yield, functional mRNA transcripts from a DNA template. This process uses purified RNA polymerase enzymes, such as T7, that recognize specific promoter sequences in the DNA template, generating the RNA transcripts of interest. However, IVT reactions also generate unwanted dsRNA byproduct. Below, we delve into some of the major quality control (QC) considerations and strategies to reduce dsRNA byproducts.
Labeled antibodies are indispensable tools in research and clinical diagnostics, used in everything from cell imaging and ELISAs to immunotherapies and ADC development. But if youโve ever tried labeling antibodies the traditional wayโpurify, buffer exchange, conjugate, purify againโyou know it can be tedious and time-consuming. Thatโs where on-bead conjugation steps in with a solution.
Tuberculosis (TB) remains one of the deadliest infectious diseases globally, with millions of new cases and over a million deaths each year. The rise of drug-resistant strains has only complicated treatment and control efforts, turning TB into a moving target for clinicians and public health officials alike. Understanding how TB spreads, evolves and becomes resistant requires more than just microscopes and culturesโit demands a detailed look at the bacteriumโs genetic code.
The new lyophilization equipment will more than double the lyophilization capacity of Promega Madison.
On March 12, 2025, a 46,000-pound stainless-steel chamber made a five-hour journey through Feynman Center to its final resting place in the brand-new Fill-Lyophilize-Finish suite. This massive piece of equipment will more than double the lyophilization capacity at Promega Madison, safeguarding the continuity of production and opening new frontiers in product formulation.
Lyophilization provides scientists with increased stability, enhanced flexibility and protection against error. Promega has been lyophilizing reagents in-house since the mid-1990s, and demand has steadily grown over time. The recent expansion reflects the companyโs commitment to anticipating scientistsโ future needs and planning for the long term.
Why is Lyophilization Important?
Lyophilization, also known as freeze-drying, provides a variety of benefits in the lab. For example, lyophilized reagents can typically be stored at higher temperatures, and they offer longer stability.
Stuart Forsyth inspects the lyophilization chamber during its installation.
โLyophilized product also gives you added flexibility in how you tailor your reagents to your specific need,โ says Stuart Forsyth, Sr Process Validation Engineer at Promega. โWhether youโre reconstituting with a buffer, water or even a sample, youโre able to alter the assayโs concentration and formulation in ways that are impossible with liquid formulations.โ
Promega also offers lyophilization for customers working with Promega to manufacture custom products. The flexibility helps many labs, especially diagnostics, ensure that the final reagent maximizes efficiency and ease of use for point-of-care applications.
โEspecially if youโre lyophilizing the whole assay in one, youโre removing a lot of potential for mistakes by the user that would result in product failure,โ says Terri McDonnell, Director of Global Custom & OEM Commercial Development. โLyophilization capabilities are powerful tools to have in your toolbox as you try to formulate a reagent for minimal risk of misuse or mistakes.โ
Expanding Lyophilization at Promega Madison
The new lyophilizer will primarily be used with 10ml vials and 100ml bottles, but it can process numerous other formats.
Promega Operations closely monitors the throughput capacity of all critical processes. For years, the team has projected that manufacturing would outgrow the existing lyophilization capacity sometime in the mid-2020s. The project to build out the empty suite in Feynman Manufacturing Center began in 2021, and it will start producing products for sale in early 2026.
The new lyophilizer nearly doubles the throughput capacity of Promega Madison. It will primarily be used with 10ml vials and 100ml bottles, but the line can also handle 2ml and 3ml vials and large LyoGuard trays for bulk powder production. At this point, the team plans to primarily use the Feynman suite for high-demand catalog products like CellTiter-Glo, creating flexibility to use the older lines for custom products and other smaller demands.
Continuity, Collaboration and Creativity
The new lyophilization suite will have several significant impacts for scientists using Promega reagents.
First, the new lyophilization line creates additional redundancy to ensure that key products are continuously available. The huge increase in capacity means that if one lyophilizer is down for maintenance, the others can handle picking up the slack. The new suite also features the current state-of-the-art automation technology, minimizing any risks for contamination or human error that would disrupt high-quality production.
The lyophilizer is unloaded by crane outside Feynman Manufacturing Center.
For customers working with Promega on custom orders, the new lyophilizer gives Promega more flexibility to collaborate with customers on finding the right formulation for their needs, all within the established quality system.
“We partner with a wide range of customers seeking to adapt or customize our technologies for specific applications,โ says Terri McDonnell. โAs the primary manufacturer of most of our products, and with the addition of new lyophilization capabilities, we can offer expanded scale and format options. Because these activities are performed in-house, we maintain greater control over quality and supply chain logistics, helping to ensure the consistent and reliable delivery of products.”
Finally, the additional capacity means that high-volume products can be manufactured less frequently by scaling up batch sizes. This frees up human resources to explore process improvements and dedicate more time to work outside of the production workflow. Kris Pearson, Director of Manufacturing Sciences and Custom operations, says the smaller equipment can serve as a sandbox where teams can test creative ideas.
โWeโll have more opportunity to work with R&D on new product development, and to dive deep into new cycles and what that can mean for our custom capabilities,โ she says. โWe can play around with new formats and processes to find new ways of offering a great product for every custom customer.โ
Long-Term Planning and Strategy
As a private company, Promega isnโt beholden to short-term gains. Leadership prioritizes decisions that support future needs, while building in room to adapt to changes in the scientific landscape.
The architectural drawings of Feynman Manufacturing Center show the suite earmarked for lyophilization as early as 2012, before the building was constructed.
โWhen we started designing Feynman Manufacturing Center, we said we wanted 30% of the square footage to be frontier space,โ says Jen Romanin VP of Global Support and IVD Operations, and key member of the Global Planning Team. โThis space would give us future flexibility in where new features would be installed.โ
Sometimes needs are forecasted far in advance โ for example, the architectural drawings of Feynman Manufacturing Center dated February 2012 show the new suite was already earmarked for Lyophilization almost a decade before the construction project began. Other spaces are left intentionally unlabeled as a nod to the unknown needs that will emerge over time. Whatever arises, the flexibility and foresight built into Promega facilities will position the team to respond quickly โ and build a high-quality solution โ without having to break new ground.
โI think this says two things about us,โ says Chuck York, Vice President of Operations at Promega. โFirst, it says weโre pretty confident weโre going to be here for a long time. Secondly, it says that no matter what happens between now and then, we want to make sure weโre prepared.โ
In recent years, non-coding RNAsโespecially microRNAs (miRNAs) and long non-coding RNAs (lncRNAs)โhave emerged as powerful regulators of cellular behavior. These molecules modulate gene expression, often by targeting mRNAs for translational suppression or degradation. Two recent studiesโone focused on osteoarthritis and the other on 5-Fu-resistant colorectal cancerโillustrate how these non-coding, regulatory RNAs operate within disease-relevant signaling networks, providing new points for therapeutic intervention.
Both studies use the pmirGLO Dual-Luciferase miRNA Target Expression Vector to evaluation predicted miRNA activity. This dual-luciferase system offers a clean and quantifiable way to validate miRNAโmRNA interactions using a simple bioluminescent readout. By cloning the 3ยด untranslated regions (UTRs) of suspected targets downstream of a firefly luciferase reporter and normalizing against Renilla luciferase, researchers can rapidly confirm whether a miRNA directly regulates its target.
Antibody-drug conjugates (ADCs) are an increasingly powerful class of cancer therapeutics that combine the targeted precision of monoclonal antibodies with the cytotoxic potency of small-molecule drugs. By directing chemotherapy agents specifically to tumor cells, ADCs aim to maximize antitumor activity while minimizing damage to healthy tissues. One key challenge in ADC design is selecting the right target and payloadโfeatures that define efficacy, safety and resistance.
The brain is constantly rewiring itself, fine-tuning connections that shape how we think, learn, and remember. But capturing those fleeting molecular changes as they happen โ at the level of individual synapses and across entire brain regions โ has long been a challenge in neuroscience. Now, thanks to recent advances in HaloTagยฎ dye technology, researchers can visualize protein dynamics in living brains with stunning clarity and specificity.
From enzyme activity to gene expression, light-based assays have become foundational tools in life science research. Among these, fluorescence and bioluminescence are two of the most widely-used approaches for detecting and quantifying biological events. Both rely on the emission of light, but the mechanisms generating that lightโand the practical implications for experimental designโare quite different.
Choosing between a fluorescence or bioluminescence assay isnโt as simple as picking between two reagents off the shelf. Each has strengths and limitations depending on the application, instrumentation, and biological system. In this blog, weโll walk through how each method works, where they shine (and where they donโt), and what to consider when deciding which approach is right for your experiment.
Todayโs blog is written by guest blogger, Kai Hillman, Associate Product Marketing Manager at Promega.
RNA therapeutics have revolutionized modern medicine, offering groundbreaking solutions for diseases that were once deemed untreatable. These innovative treatments harness the power of RNA molecules to correct genetic anomalies and modulate protein expression, paving the way for personalized medicine. Among the many facets of RNA biology, double-stranded RNA (dsRNA) plays a pivotal role in cellular processes and immune surveillance.
XWe use cookies and similar technologies to make our website work, run analytics, improve our website, and show you personalized content and advertising. Some of these cookies are essential for our website to work. For others, we wonโt set them unless you accept them. To learn more about our approach to Privacy we invite you to Read More
By clicking โAccept Allโ, you consent to the use of ALL the cookies. However you may visit Cookie Settings to provide a controlled consent.
We use cookies and similar technologies to make our website work, run analytics, improve our website, and show you personalized content and advertising. Some of these cookies are essential for our website to work. For others, we wonโt set them unless you accept them. To find out more about cookies and how to manage cookies, read ourย Cookie Policy.
If you are located in the EEA, the United Kingdom, or Switzerland, you can change your settings at any time by clickingย Manage Cookie Consentย in the footer of our website.
Necessary cookies are absolutely essential for the website to function properly. These cookies ensure basic functionalities and security features of the website, anonymously.
Cookie
Duration
Description
cookielawinfo-checbox-analytics
11 months
This cookie is set by GDPR Cookie Consent plugin. The cookie is used to store the user consent for the cookies in the category "Analytics".
cookielawinfo-checbox-functional
11 months
The cookie is set by GDPR cookie consent to record the user consent for the cookies in the category "Functional".
cookielawinfo-checbox-others
11 months
This cookie is set by GDPR Cookie Consent plugin. The cookie is used to store the user consent for the cookies in the category "Other.
cookielawinfo-checkbox-advertisement
1 year
The cookie is set by GDPR cookie consent to record the user consent for the cookies in the category "Advertisement".
cookielawinfo-checkbox-necessary
11 months
This cookie is set by GDPR Cookie Consent plugin. The cookies is used to store the user consent for the cookies in the category "Necessary".
cookielawinfo-checkbox-performance
11 months
This cookie is set by GDPR Cookie Consent plugin. The cookie is used to store the user consent for the cookies in the category "Performance".
gdpr_status
6 months 2 days
This cookie is set by the provider Media.net. This cookie is used to check the status whether the user has accepted the cookie consent box. It also helps in not showing the cookie consent box upon re-entry to the website.
lang
This cookie is used to store the language preferences of a user to serve up content in that stored language the next time user visit the website.
viewed_cookie_policy
11 months
The cookie is set by the GDPR Cookie Consent plugin and is used to store whether or not user has consented to the use of cookies. It does not store any personal data.
Analytical cookies are used to understand how visitors interact with the website. These cookies help provide information on metrics the number of visitors, bounce rate, traffic source, etc.
Cookie
Duration
Description
SC_ANALYTICS_GLOBAL_COOKIE
10 years
This cookie is associated with Sitecore content and personalization. This cookie is used to identify the repeat visit from a single user. Sitecore will send a persistent session cookie to the web client.
vuid
2 years
This domain of this cookie is owned by Vimeo. This cookie is used by vimeo to collect tracking information. It sets a unique ID to embed videos to the website.
WMF-Last-Access
1 month 18 hours 24 minutes
This cookie is used to calculate unique devices accessing the website.
_ga
2 years
This cookie is installed by Google Analytics. The cookie is used to calculate visitor, session, campaign data and keep track of site usage for the site's analytics report. The cookies store information anonymously and assign a randomly generated number to identify unique visitors.
_gid
1 day
This cookie is installed by Google Analytics. The cookie is used to store information of how visitors use a website and helps in creating an analytics report of how the website is doing. The data collected including the number visitors, the source where they have come from, and the pages visted in an anonymous form.
Advertisement cookies are used to provide visitors with relevant ads and marketing campaigns. These cookies track visitors across websites and collect information to provide customized ads.
Cookie
Duration
Description
IDE
1 year 24 days
Used by Google DoubleClick and stores information about how the user uses the website and any other advertisement before visiting the website. This is used to present users with ads that are relevant to them according to the user profile.
test_cookie
15 minutes
This cookie is set by doubleclick.net. The purpose of the cookie is to determine if the user's browser supports cookies.
VISITOR_INFO1_LIVE
5 months 27 days
This cookie is set by Youtube. Used to track the information of the embedded YouTube videos on a website.
Performance cookies are used to understand and analyze the key performance indexes of the website which helps in delivering a better user experience for the visitors.
Cookie
Duration
Description
YSC
session
This cookies is set by Youtube and is used to track the views of embedded videos.
_gat_UA-62336821-1
1 minute
This is a pattern type cookie set by Google Analytics, where the pattern element on the name contains the unique identity number of the account or website it relates to. It appears to be a variation of the _gat cookie which is used to limit the amount of data recorded by Google on high traffic volume websites.