BioSpring continues to grow: new capacities for genome editing applications

More flexibility, more speed - as the global market leader for active pharmaceutical ingredients for genome editing applications, our clients will have access to significantly expanded production capacities to meet growing demand. New state-of-the-art production facilities have been built at our second site in Frankfurt, Germany. In the four new cleanrooms, we produce mRNA and guide RNA tailored for therapeutic genome editing applications. In additional new laboratories for mRNA research and analysis, we conduct process development and optimization of small-scale non-GMP projects. 

The new cleanrooms with system-controlled simultaneous production of Good Manufacturing Practice (cGMP) guide RNA allow processing of several client orders at the same time. In this way our clients benefit from improved turnaround times and enhanced flexibility for their clinical applications.

With the upcoming cGMP certification for the therapeutic production of mRNA, our clients will be able to source both cGMP-certified guide RNA and mRNA for their genome editing applications from us as their sole supplier. With the new production facility, we are tripling our manufacturing capacity for guide RNA, enabling us to produce over 40 additional batches per year. Already today, most clinical guide RNA material worldwide is manufactured at BioSpring facilities.

To sustain our growth trajectory, we are developing additional large-scale production facilities, for which we have already acquired a 40,000 square meters industrial site in Offenbach am Main. This is positioning us for continued expansion and innovation in the future.

At this year's TIDES US conference, Dr. Barbara Karolina Pfaff, QC Manager of Molecular Sequencing, delivered an insightful presentation on the transformative potential of Next-Generation Sequencing (NGS) in oligo-analytics. Her talk highlighted how NGS technology is revolutionizing the field, offering unprecedented precision and depth in oligonucleotide analysis.

Benefits for our clients:

• Detailed impurity characterization: enables detection of impurities and sequence variants, providing deeper insights into molecular composition
• Accurate results: relies on precise library preparation, ensuring reliable sequencing data
• Innovative bioinformatic tools: our tool OligoXplore enhances data analysis, helping identify impurities and sequence variants with high precision
• Customized data analysis pipelines: tailored solutions support advanced research and therapeutic development
• Future potential: advanced NGS techniques promise new possibilities for therapeutic development and molecular research

NGS is vital for oligo-analytics, particularly in meeting FDA recommendations for guide RNA purity analysis. Its sensitivity and accuracy are crucial for ensuring the purity and efficacy of therapeutic oligonucleotides, aligning with stringent regulatory standards. Furthermore, NGS enables detailed impurity characterization and sequence variant detection, providing researchers with deeper insights into the molecular composition and potential modifications or impurities of oligonucleotides.

Accurate NGS results depend on precise library preparation. The Illumina library preparation process for guide RNA involves critical steps that ensure reliable and reproducible sequencing data. This foundational aspect of NGS is essential for subsequent data analysis and interpretation, underscoring the importance of meticulous library preparation. At BioSpring, we introduced OligoXplore, which is a tailored bioinformatic solution designed specifically for NGS data analysis. This innovative tool facilitates the interpretation of complex sequencing data, helping researchers identify impurities and sequence variants with high precision. Effective NGS data analysis is essential for deriving meaningful insights from sequencing projects, making tools like OligoXplore invaluable.

Customized data analysis pipelines enhance the utility of NGS, providing researchers with the tools they need to address specific research challenges. These tailored bioinformatic solutions at BioSpring offer unparalleled insights into oligonucleotide purity and functionality, supporting advanced research and therapeutic development.

Dr. Pfaff's presentation at TIDES US underscored the immense potential of NGS in oligonucleotide research. Through advanced NGS techniques and bioinformatic tools, researchers can achieve a level of detail and accuracy previously unattainable. This transformative technology promises new possibilities for therapeutic development and molecular research.

Want to know more? Download our poster on NGS right here. 

At this year’s CRISPR Medicine Conference and at TIDES US, Dr. Raoul Hennig, Head of Manufacturing Site II, delivered a captivating presentation on the evolution and efficiency of guide RNA manufacturing at BioSpring. His in-depth analysis offered attendees a comprehensive overview of our journey towards becoming the market leader in guide RNA production, focusing on driving efficiency, process scalability, and future innovations.

Benefits for our clients:

• Enhanced efficiency and robustness of our guide RNA manufacturing process: optimized manufacturing processes ensure high quality and reliability for gene-editing technologies like CRISPR/Cas
• Continuous improvement: we continuously work on improving our manufacturing processes to maintain market leadership position
• Process scalability: we ensure process scalability to meet the increasing demand for guide RNA while maintaining highest quality standards
• Future initiatives: we are working on further refining processes and driving innovation in guide RNA manufacturing

Over the past few years, BioSpring has made significant strides in optimizing its manufacturing processes, which have been instrumental in solidifying our leadership position. These advancements have not only enhanced the efficiency and robustness of the platform process but also ensured the high quality and reliability essential for the success of gene-editing technologies like CRISPR/Cas.

BioSpring's relentless focus on continuous improvement has led to systematic advancements from 2016 to 2020, transforming the manufacturing workflow. The introduction of Process 2.0 marked a pivotal upgrade, which translated into more consistent and reliable gRNA production. This evolution has been crucial for maintaining the high standards required in the industry.

A key highlight in recent advancements has been BioSpring's approach to process scalability. We have successfully scaled our manufacturing processes to meet the increasing demand for gudie RNA across various applications, all while maintaining the highest quality standards. This achievement underscores BioSpring's commitment to providing reliable and scalable solutions to our clients.

Our strategic vision since last year focuses on sustaining efficiency gains and further refining manufacturing processes to adapt to new challenges and opportunities. This forward-thinking approach ensures continued leadership in guide RNA manufacturing. The roadmap ahead for BioSpring is filled with exciting projects aimed at pushing the boundaries of what’s possible in guide RNA manufacturing. Future initiatives are designed to drive further efficiencies and innovations, ensuring access for our clients to the most advanced manufacturing and analytics technologies.

The comprehensive overview of the evolution and efficiency of BioSpring's guide RNA manufacturing process highlights a journey of continuous improvement and innovation. As BioSpring continues to drive the future of guide RNA manufacturing, it remains dedicated to providing updates on progress and innovations, promising an exciting horizon ahead.

Great success for our analytical research department! In a highly acclaimed scientific paper, Dr. Michael Rühl and his team have described a new method developed in-house for the difficult challenge of sequencing long oligonucleotides. This means that even large DNA molecules can now be prepared in such a way that their sequence can be confirmed using mass spectrometry, increasing the sequence coverage of the oligonucleotides from approximately 15-25% to 100%.

Benefits for our clients:

• Increased sequence coverage: up to 100% sequence coverage for long oligonucleotides, ensuring thorough verification
• Enhanced quality control: robust method for quality control in regulated environments, crucial for drug development
• Reliability: reliable technique for confirming sequences of synthetic DNA, aiding the development of therapeutic oligonucleotides
• Versatility: applicable to range of DNA molecules, supporting various research and development needs
• Regulatory compliance: aligns with stringent regulatory standards

Introduction:

The importance of oligonucleotide therapeutics, such as antisense oligonucleotides (ASOs) and small interfering RNAs (siRNAs), has grown significantly, particularly for treating inherited diseases. Technologies like CRISPR-Cas9 have also highlighted the need for accurate DNA sequencing. While mass spectrometry (MS) is a standard method for analyzing RNA therapeutics, its application to long DNA molecules has been limited due to the challenges in sequencing accuracy beyond 60 nucleotides.

Methodology:

We developed a method that combines restriction enzyme digestion with high-resolution tandem mass spectrometry. The process begins with the use of common restriction enzymes to create specific digestion products from DNA molecules. These products are then analyzed using ion pair reverse phase chromatography coupled to a time-of-flight mass spectrometer. This setup allows for collision-induced dissociation (CID) for sequence analysis.

Results:

The study demonstrated that using restriction enzymes significantly increased sequence coverage from 23.3% (using direct CID-MS/MS) to 100%. The approach involved generating partial duplexes with enzyme-directing oligonucleotides, enabling site-specific cleavage by the restriction enzymes. The resulting smaller DNA fragments were easier to analyze, allowing for complete sequence verification. This method was successfully applied to single-stranded DNA molecules and a double-stranded 131 base pair DNA duplex, confirming the sequences with high accuracy.

Discussion:

We highlight the method's applicability for research, development, and quality control in regulated environments. This approach is especially valuable for confirming the sequences of synthetic DNA molecules used in therapeutics. The study notes that while traditional methods like Sanger sequencing and next-generation sequencing (NGS) are widely used, they often cannot confirm the sequence of the target molecule directly or detect certain modifications. The restriction enzyme mediated approach provides an additional layer of sequence confirmation, ensuring the highest standards of accuracy.

Conclusion:

This novel method offers a reliable and efficient solution for confirming the sequences of long synthetic DNA molecules, crucial for the development of DNA-based therapeutics. The approach's ability to provide 100% sequence coverage and its suitability for quality control makes it a valuable tool in the pharmaceutical industry. Future work may explore the applicability of this method to synthetic DNA with chemical modifications and other complex DNA structures, further enhancing its utility in genetic research and therapeutic development.

BioSpring and Pantherna agree on strategic cooperation in the field of genome editing

The focus of our collaboration targets further development and licensing of therapeutic mRNA molecules based on PTXmRNA® technology specifically for genome editing applications. BioSpring is the world's leading provider of RNA for genome editing applications. Pantherna specializes in the development of novel lipid nanoparticle (LNP) formulations (PTXDLNP® technology) for the organ- and cell-selective uptake of therapeutically active mRNA (PTXmRNA®) for the effective expression of proteins in various diseases.

"With this research collaboration, we are combining the outstanding expertise of our companies in the field of therapeutic mRNA, creating synergies and thus developing the potential for innovative solutions for our clients," stated Dr Hüseyin Aygün, Chief Scientific Officer (CSO) and co-owner of BioSpring GmbH.

Dr Jörg Kaufmann, CSO of Pantherna Therapeutics GmbH, adds:  "We are pleased to support Biospring and its customers seeking solutions for customized mRNA constructs and nucleic acid delivery systems in the field of genome editing with our mRNA-LNP expertise."

With this new collaboration, we are placing particular emphasis on utilizing mRNA technology within the rapidly expanding field of genome editing.