Our company will be exhibiting at the 28th Annual Meeting of the Japanese Society of Gene and Cell Therapy, which will be held from Thursday, July 14. At our booth, you can see devices such as the Stunner, which can measure AAV aggregation and Full/Empty ratios in high throughput, and the Uncle, which evaluates thermal stability such as aggregation, as well as videos and materials. We look forward to your visit.

**Seminar Title** An Innovative Platform to Accelerate the Stability Evaluation of AAV Vectors **Overview** AAV is one of the viruses used in the development and basic research of gene therapy, but its characterization techniques require many considerations. In particular, stability evaluation is very important for ensuring safety. In this seminar, we will introduce a low-volume, high-throughput platform that accelerates stability evaluation, using AAV as an example.

Viscosity data is essential for the formulation, characterization, and quality control of biopharmaceuticals; however, most viscosity measurements require large amounts of samples and extensive working hours, making it difficult to obtain sufficient viscosity data. In this seminar, we will review actual measurement results of antibodies and other proteins using the high-speed micro-volume viscometer, Honeybun, designed for biological formulations, and delve into its excellent precision, accuracy, and throughput from low-volume samples.

Our company will be exhibiting at Bio Japan 2022, which will be held from Wednesday, October 12. At our booth, you can view technical data and videos of our protein stability evaluation device, Stunner, as well as new products such as the high-speed viscometer Honeybun and the fully automated buffer exchange device UNAGI. Additionally, visitors to our booth will receive original Unchained Labs merchandise as a gift! We look forward to seeing you there.

Long-term stability testing is very important for understanding the stability of biological products. However, traditional long-term stability tests require many samples and time, and one must wait until the final stages of development. The thermal stability evaluation platform tool Uncle has expanded to include isothermal change measurements in addition to Tm and Tagg measurements, allowing for the differentiation and evaluation of samples that could not be distinguished by traditional thermal stability assessments. In this webinar, we will introduce a method for conducting stability tests using Uncle that is simple, requires only a small amount of sample, and can be performed at an early stage of development.

Creating perfect lipid nanoparticles (LNPs) requires technology. Testing flow rates and formulations one by one to find the appropriate size of LNPs is time-consuming and wastes valuable payload. Additionally, scaling up often requires new workflows, and there are always such challenges in new gene therapy research. Nunchuck is a groundbreaking platform that combines two critical steps necessary for producing LNPs of optimal size: Total Flow Rate (TFR) screening and LNP formulation exploration. It can screen eight types of TFRs in just 10 minutes, quickly finding the sweet spot and allowing immediate production of LNPs with capacities ranging from 0.5 to 200 mL. Moreover, the produced LNPs can be quickly evaluated for physical properties, such as payload measurement, using Stunner. This webinar will introduce how to conduct TFR screening suitable for the purpose of creating size-tunable, high-quality RNA-LNPs and how to scale up. We will also demonstrate how to evaluate the produced LNPs with Stunner.

Our company will be exhibiting at the 29th Annual Meeting of the Japanese Society of Gene and Cell Therapy, which will be held from September 11 (Monday) to September 13 (Wednesday), 2023. At our booth, we will introduce groundbreaking solutions for gene therapy research and development, including the stability evaluation device Stunner and our new product Leprechaun. We invite you to visit our booth.

This seminar is jointly hosted by Kitahama Seisakusho Co., Ltd. and Unchained Labs, Inc. In the field of LNP creation technology, we will introduce a technique for producing monodisperse nanoparticles such as LNPs using the microfluidic devices from ParticleWorks (formerly Blacktrace), which became a brand of Unchained Labs in July of this year. Additionally, we will present various automation devices that leverage Unchained Labs' strengths in biopharmaceutical research and gene therapy in the evaluation and post-processing technology, offering solutions to the challenges faced by researchers in biopharmaceutical analysis and gene therapy drug development through these products. <Presentation 1> Streamlining LNP formulation development using microfluidic technology Moriho Fukai, Kitahama Seisakusho Co., Ltd. <Presentation 2> A tool that significantly enhances the efficiency of buffer exchange and characterization of biopharmaceutical samples using lipid nanoparticles Yuji Senoo, Unchained Labs, Inc.

Lipid nanoparticles (LNPs) are currently one of the most notable delivery systems used in mRNA vaccines, gene therapy, cancer treatment, and more. However, their development requires a complex process involving formulation screening, control of size and quality, removal of organic solvents, and characterization, which demands significant effort and time. In this webinar, we will introduce methods to enhance the production of perfectly sized LNPs, characterization of size and quality, buffer exchange and sample concentration, and stability analysis techniques at each step of the development process workflow.

Topic: In recent years, viruses have been widely used as modalities for gene therapy and vaccines, and research and development have been advancing. Adenoviruses are one of the representative viruses, but traditional evaluation methods are diverse and require a lot of time and effort. For example, obtaining quick answers regarding the capsid titer, Full/Empty ratio, and the presence of aggregation is a significant challenge. The UV/Vis absorbance at 260nm (A260) is considered the fastest method to confirm the titer of adenoviruses, but it requires denaturing the virus, resulting in an assay time of about 30 minutes. Additionally, the results of A260 can be inhibited by contamination from proteins, DNA, and various other substances. Furthermore, titer measurement methods like qPCR are not only time-consuming and labor-intensive but also consume a large amount of samples, making them unsuitable for rapid checks. Seminar Overview: In this seminar, we will explain how Stunner can quickly and accurately solve these challenges as a one-stop solution. We will also look at the application of adenovirus vector sample QC and the evaluation of various types of adenoviruses.

CPhI Japan Exhibition & Mini Seminar! April 19 (Wed) - April 21 (Fri) @ Tokyo Big Sight Unchained Labs, Inc. will be exhibiting in the biopharmaceutical zone at CPhI Japan this year as well. This time, we will also hold a mini seminar at our booth with plenty of participation benefits! Please come visit our booth at Tokyo Big Sight. <Mini Seminar> - "Mastering the Research and Development of Lipid Nanoparticles (LNP) to Earn Your Black Belt" - "Let Automation Handle Buffer Exchange and Go Home on Time" *Please check the [Pre-registration Form] for the schedule. <Mini Seminar Participation Benefits> [Benefit 1] Pre-register to receive our original eco-bag that fits a PC! [Benefit 2] Attend on the day to gain a chance to win a 3000 yen QUO card! [Benefit 3] Get technical questions and consultations answered on the spot. <Pre-registration Form> Register now using the link below to get [Benefit 1]! *Separate web pre-registration or invitation is required for entry to the exhibition.

Topic: The development of biopharmaceuticals is becoming mainstream, but many challenges remain due to their complex structures and properties compared to small molecule drugs. For example, long-term stability testing of proteins in biopharmaceuticals is generally conducted in the later stages of biopharmaceutical development. This is because it requires a large amount of samples and time, making it unsuitable for early candidate drug screening. However, this approach carries the risk that issues with candidate drugs may only be discovered in the later stages of development. Additionally, controlling the viscosity of formulations is also an important development requirement, yet obtaining viscosity data can take too much time. Traditional methods require a large number of samples and can only measure one sample at a time. Even if there is a technology that can measure with just a few µL of sample, it still requires significant maintenance and time for tasks such as clog removal and cleaning. Webinar Overview: In this webinar, we will tackle these challenges in biopharmaceutical development and introduce methods for conducting isothermal stability testing in the early stages of development, as well as a technique for simultaneously measuring the viscosity of multiple samples in just a few minutes without the need for cleaning.

ADC (antibody-drug conjugate) is a biopharmaceutical that is attracting attention due to its pharmacological activity and the excellent specificity of antibodies. However, the characterization of ADCs is very challenging due to the rarity of samples and their tendency to aggregate. In this webinar, we will explain how to obtain important information such as the concentration, labeling, stability, and viscosity of mAbs and their conjugates from just a few microliters of samples, along with actual data to demonstrate the evaluation of ADC characteristics.

<Announcement of Interphex Exhibition and Product Technology Seminar> Our company will be exhibiting at the Interphex Pharma Lab EXPO again this year. Additionally, on the first day of the exhibition, July 5th, we will also be presenting at a product technology seminar! We invite you to come to Tokyo Big Sight. Exhibition Details: Name: Interphex Week Tokyo (25th Edition) Date: July 5 (Wednesday) - July 7 (Friday) 10:00-17:00 Venue: Tokyo Big Sight, East Hall 1, Booth: 1A-10 Details of our exhibited products: https://www.interphex.jp/tokyo/ja-jp/search/2023/directory/directory-details.org-fc5952a0-dbf6-4ef7-bfa0-2feb60a9169f.html Product Technology Seminar: "Innovative Next Steps in Pharmaceutical Development Using Lipid Nanoparticles" We will introduce a solution that allows for high-throughput processes from the examination of conditions for lipid nanoparticle production to buffer exchange and physical property evaluation. Date and Time: July 5 (Wednesday) 12:30-13:00 Venue: Room 4 (East Hall 5)

Our company will be exhibiting at the 23rd Annual Meeting of the Protein Science Society, which will be held from July 5 (Wednesday) to July 7 (Friday), 2023. At our booth, we will introduce groundbreaking solutions for the research and development of proteins and antibody drugs, including our characterization device, Stunner. We invite you to visit our booth. Dates: July 5 (Wednesday) - July 7 (Friday), 2023 Venue: Nagoya Congress Center Booth: #18 Details: https://www2.aeplan.co.jp/pssj2023/

Challenge: Monitoring the titer, structural stability, and non-viral contaminants of lentiviruses during production is crucial for generating high-yield and high-purity pharmaceuticals. However, traditional methods such as ELISA and qPCR require sample purification and cannot perform structural analysis of the virus, making it very difficult to obtain accurate viral titers that are not affected by fragmented viral particles or contaminants. Solution: Unchained Labs' new product, Leprechaun, is a platform that provides critical information about crude or purified lentivirus samples. Leprechaun measures particle size to separate individual lentiviruses from aggregates and uses fluorescent antibodies to check for the presence of VSVG, capsid, soluble p24, and contaminants from extracellular vesicles (EVs). By using Leprechaun, it is possible to monitor titer and purity throughout the entire workflow of lentivirus research and development. In this webinar, we will explain how Leprechaun reveals the truths about your lentivirus that you have never known before.

In the evaluation of AAV characteristics, there is always a demand for rapid responses regarding measurement criteria such as capsid titer, genome titer, Empty/Full ratio, aggregation, total protein amount, and DNA amount. By combining this data, we can understand what is happening during sample and process development. However, since there are many characterization analysis methods available, the key to success lies in how to select appropriate analytical tools and how much data can be interpreted from rare samples. In this webinar, we will explain how our Stunner technology can assist in rapid sample QC, optimization of downstream purification chromatography, and improvement of workflows for other common analytical assays. We will present case studies from actual paper data demonstrating how Stunner accelerates AAV characterization and saves both samples and time.

In conventional protein quantification methods, one had to choose between high precision and high throughput. Plate-based UV/Vis measurements can provide instant readings, but their accuracy and precision are not very high. On the other hand, other measurement methods can achieve high precision, but they are time-consuming. Using our Lunatic for protein quantification offers superior precision, accuracy, and throughput compared to other methods, allowing for the measurement of 96 samples in 10 minutes with an accuracy within 2%. Additionally, Lunatic requires only 2μL per sample, making it very efficient. In this webinar, you will see data on the precision, reproducibility, and throughput of protein quantification using Lunatic. Even tricky samples that do not perform well with other UV/Vis readers can be analyzed without issues, as Lunatic's Unmix technology can identify contamination. Furthermore, we will also explain how to perform QC checks with the addition of DLS (Dynamic Light Scattering) technology.

Our company will be exhibiting at BioJapan 2023, which will be held from Wednesday, October 11. At our booth, you can see the protein structural stability evaluation device, Uncle, as well as technical data on Leprechaun, which can analyze lentiviruses and exosomes, and a mysterious new product. Additionally, on the first day, we will have a lecture by Professor Nagamori from the University of Tokyo on the broad drug discovery support provided by Uncle. We invite you to come to Pacifico Yokohama! Exhibition details: ■ Dates: October 11 (Wednesday) to October 13 (Friday), 2023 ■ Venue: Pacifico Yokohama ■ Booth #: B-36 Seminar details: "Evaluation methods for physical properties enabling broad drug discovery support from small molecule drugs to antibody drugs" ■ Date: October 11 (Wednesday) ■ Time: 14:30 - 15:30 ■ Location: Annex Hall F203 ■ Details and pre-registration: https://biojapan2023.jcdbizmatch.jp/Lookup/jp/Seminar/u0?ke=unchained

Lipid nanoparticles (LNPs) are used as carriers to deliver active ingredients such as mRNA into cells. However, their development involves many optimization factors in terms of the formulation of components (lipids, payload) and manufacturing conditions. Therefore, optimizing requires a large number of experiments and comparative evaluations, which is considered to be a tremendous effort and time-consuming. In this webinar, we will introduce our solutions that can address this challenge. First, we will explain the Sun series of LNP manufacturing devices, which can seamlessly execute and transition from efficient formulation and manufacturing condition screening to GMP-compliant production. We will also introduce the Stunner analysis device, which provides various information such as LNP size and concentration at high throughput, and the Unagi automatic buffer exchange device, which aids in solvent removal and formulation optimization, further accelerating development.

Unchained Labs, Inc. will be exhibiting at the 3rd Annual Meeting of the Japanese Society for Antibody Research, which will be held from December 9 (Monday). At our booth, we will introduce innovative tools essential for antibody and ADC research, including the Uncle device for protein structural stability evaluation and the Stunner, which can measure various parameters such as potency and size. Additionally, on the 10th, Professor Akira Nagamori from the University of Tokyo will give a lecture on physical property evaluation using our Uncle device during a luncheon seminar. We look forward to seeing you in Sendai! Exhibition details: ■ Dates: December 9 (Monday) to 11 (Wednesday), 2024 ■ Venue: Sendai International Center ■ Booth #: 38 Luncheon seminar details: "Capturing Antibodies and Small Molecules with the Physical Property Evaluation Device Uncle!" ■ Speaker: Professor Akira Nagamori, Graduate School of Engineering, University of Tokyo ■ Date: December 10 (Tuesday) ■ Time: 12:15 PM - 1:15 PM ■ Location: Sendai International Center Exhibition Hall, Satellite Room 2 (Conference Room 2)