What if the bottleneck in cell line development isn’t how many clones you screen, but how you track them?

Cloning workflows have long relied on brute force: screen more cells, automate harder, and hope that small-scale performance predicts manufacturability. But too often, the “perfect” clone in a 96-well plate turns into a dud when it reaches the bioreactor. That disconnect costs time, money, and promising therapies.

This week, host David Brühlmann welcomes Kent Rapp, Co-founder and CEO of Biolinco, an entrepreneur who’s turning the classic approach to cell line development inside out. Drawing from his background in chemical engineering and his work in biomanufacturing at Johns Hopkins University, Kent teamed up with DNA barcoding experts to pioneer a new workflow: barcode every cell, pool them, and track their true performance in the environment that matters.

Topics discussed:

• The pitfalls of brute-force screening in traditional cell line development (03:05)
• Kent’s background and how he was drawn to combine science, startups, and biomanufacturing (04:37)
• Overcoming discrepancies between small-scale and large-scale screening environments (08:30)
• How DNA barcoding allows for high-resolution, pooled clone screening (10:34)
• Sensitivity advantages of sequencing over plate-based detection (14:21)
• Methodology for tracking and recovering individual high-performing clones from pools (15:13)
• Impact on speed and workflow efficiency in cell line development (17:31)
• Regulatory and safety considerations related to DNA barcodes in cell lines (19:04)

Smart insight: According to Kent, biotech as an industry has a tendency to "automate problems instead of solve them". Rather than addressing the root causes—like lack of meaningful measurements at relevant scales—companies often throw more robots and more plates at the issue, hoping brute force will finally yield the magical clone. But real process improvement requires a rethinking of what is being measured and how those insights are generated—not just a higher throughput of the same flawed assay.

If this got you rethinking how you screen clones, you'll want these next. We've tackled cell line development, high-throughput screening, and the art of spotting manufacturable candidates early from a few different directions — here are four worth queuing up.

• Episodes 117 - 118 : Cell Line Development Secrets: Eliminating Critical Bottlenecks for Faster Timelines with Andrea Gough
• Episodes 09 - 10: Revolutionizing Cell-Line Development: Unleashing the Power of Nanopens and Microenvironments with Tanner Nevill
• Episodes 123 - 124: Manufacturability: Why Most Protein Candidates Fail (And How to Pick Winners Early) with Susan Sharfstein
• Episodes 115 - 116: Revolutionizing Biologics Development with Hyper Throughput Screening and AI with Jeremy Agresti

Connect with Kent Rapp:
LinkedIn: www.linkedin.com/in/kent-rapp
Biolinco website: www.biolinco.com

Free 5-day email course, The CMC Failure Chain: the five recurring CMC mistakes that put your promising program at risk → Get it here

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Your active ingredient is the nucleic acid. So why does a proteolipid vehicle filing include viral clearance studies, stability data and full characterisation of a membrane protein that is not the drug? Because that protein sits on the particle surface, and a component nobody has filed before is the agency's problem regardless of what you call it.

Proteolipid vehicles (PLVs), the platform Jitendra Kumar works on as Lead Scientist for Chemistry and Process Development at Entos Pharmaceuticals, represent a novel frontier in drug delivery, Instead of being taken up into an endosome and having to escape it, a PLV fuses with the cell membrane and releases cargo straight into the cytosol. That opens targets and patient groups that liver-tropic lipid nanoparticles and viral vectors have struggled to reach, and it leaves Kumar building a regulatory file with nothing on the shelf to copy.

Highlights from the episode:

• Strategies for communicating novel technology with regulatory agencies and ensuring robust science-driven submissions (02:34)
• What differentiates the analytical characterization of PLVs compared to standard recombinant proteins or antibodies (04:03)
• The development plan and regulatory pathway towards clinical and commercial approval for their lead leptin therapy (05:35)
• The evolving role of advanced techniques—such as cryo-EM—in supporting regulatory filings and product understanding (06:56)
• Jitendra Kumar's career journey: from agricultural research in India to protein science and neurodegeneration, and how these experiences inform current PLV technology development (09:00)
• Challenges of early diagnostics and product development in neurodegenerative diseases (14:29)
• Decision-making differences and focus in academic versus industry biotech research (15:32)
• Practical advice on the importance of honest technology assessment, building networks, and understanding both strengths and weaknesses (16:16)

Smart insight: A persistent challenge for groundbreaking delivery systems is the lack of established regulatory playbooks. Jitendra Kumar laid out a science-first approach: let data do the talking, supported by rigorous GLP toxicology studies and transparent communication with agencies. Regulatory bodies like Health Canada are receptive to innovation, provided that sponsors demonstrate safety, efficacy, and scientific rationale for any deviation from standard criteria.

If this conversation got you thinking about how novel delivery vehicles reach the cell — and what it takes to carry one from bench through CMC, scale-up, and regulatory review — these four episodes go deeper:

• Episodes 125 - 126: How to Enhance Cell Engineering Using Mechanical Intracellular Delivery with Armon Sharei
• Episodes 231 - 232: From IND to BLA: The Biologics CMC Decisions That Determine Regulatory Success with Henri Kornmann
• Episodes 89 - 90: Scale-Up Secrets: Cracking the Code of AAV Production with Ahmed Youssef
• Episodes 71 - 72: Effective Outsourcing: How Small Biotech Companies Can Thrive with Mark Melville

Connect with Jitendra Kumar:

• LinkedIn: www.linkedin.com/in/jkumar2
• Email: jitendra.kumar@entospharma.com
• Website: www.entospharma.com

Free 5-day email course, The CMC Failure Chain: the five recurring CMC mistakes that put your promising program at risk → Get it here

Support the show

Gene therapy only works if the cargo reaches the right cells intact. Adeno-associated viruses (AAV) and lipid nanoparticles have carried the field this far, but both share a constraint: the particle is taken up into an endosome, and the payload has to escape that compartment before it is degraded. Endosomal escape is where a large share of the dose is lost, and it is why delivery, not the genetic construct, is usually the thing that limits the therapy. Lipid nanoparticles carry a second constraint, since they tend to accumulate in the liver, which narrows the diseases they can reach. What if the particle never entered that way at all?

Jitendra Kumar, Lead Scientist for Chemistry and Process Development at Entos Pharmaceuticals, works on a platform that fuses directly with the cell membrane and releases its cargo straight into the cytosol. He came to nanoparticle design the long way, through fifteen years of structural biology on the prion protein in Frankfurt and Edmonton, which is why he thinks about particle size, packaging and diffusion the way he does.

Key topics discussed:

• Jitendra’s career background in structural biology and journey to Entos Pharmaceuticals (03:18)
• The scientific motivation and challenges of working with prion proteins and breaking down complex diseases (04:55)
• Genetic medicine approaches: gain-of-function vs. loss-of-function, and the role of siRNA, ASOs, and gene delivery (06:27)
• The FAST protein platform: origins, function, and advantages for drug delivery (07:56)
• Manufacturing differences compared to LNPs, including the introduction of recombinant membrane protein production and related CMC complexity (09:55)
• Scale-up and production challenges for membrane proteins, and strategies for clinical supply (11:21)
• Clinical development progress: Phase 1/2 studies with the platform, especially for COVID vaccine delivery (12:23)
• Focus areas for the technology, including selective lung delivery and leptin therapy for lipodystrophy (13:12)
• Lessons for small biotech companies in phase 1/2 manufacturing strategy, technology transfer, and the value of an experienced network (14:31)
• Balancing process robustness with speed in new biotech ventures (16:02)
• The importance of identifying “pause steps” and must-have vs. nice-to-have features in early manufacturing processes (17:15)

Smart insight: Jitendra’s takeaway for startups: for early phases, find a partner who genuinely understands your tech and can move at your pace rather than defaulting to a big CDMO, treat your network as infrastructure, and build the ability to run production in-house. The one thing that's never up for negotiation is process robustness. The real question isn't robustness vs. speed, but how many checkpoints and safe pause points you build in so you can have both.

If this conversation got you thinking about how novel delivery vehicles reach the cell — and what it takes to carry one from bench through CMC, scale-up, and regulatory review — these four episodes go deeper:

• Episodes 125 - 126: How to Enhance Cell Engineering Using Mechanical Intracellular Delivery with Armon Sharei
• Episodes 231 - 232: From IND to BLA: The Biologics CMC Decisions That Determine Regulatory Success with Henri Kornmann
• Episodes 89 - 90: Scale-Up Secrets: Cracking the Code of AAV Production with Ahmed Youssef
• Episodes 71 - 72: Effective Outsourcing: How Small Biotech Companies Can Thrive with Mark Melville

Connect with Jitendra Kumar:

• LinkedIn: www.linkedin.com/in/jkumar2
• Email: jitendra.kumar@entospharma.com
• Website: www.entospharma.com

Free 5-day email course, The CMC Failure Chain: the five recurring CMC mistakes that put your promising program at risk → Get it here

Support the show