Search bioRxiv⌕ Search

Biology subjects

Donatelli, J.

Publications and source records attributed to Donatelli, J..

3 recordsLinked to original sources

An AI/ML-Powered Workflow for End-to-End Cell Line Development

The generation of clonal CHO cell lines is foundational to biologics manufacturing; however, labor-intensive cell culture workflows predominate in the field. We created the CLAIRE (Cell Line AI Recognition and Evaluation) tool to streamline end-to-end cell line development by integrating deep-learning image analysis with automated liquid handling. We benchmarked three object detection models for monoclonality verification and found DETR provides superior accuracy (>0.90 F1-score) in identifying single cells. To quantify the outgrowth of cell lines, we evaluated multiple zero-shot SAM2 segmentation models against a feature-based estimation method. Feature-based detection successfully identified diverse cell colony types while less robust performance was observed for SAM2 models, particularly for sparse density colonies. The pre-trained DETR and feature-based detection models were wrapped in a task-focused user interface that outputs cell line hitpick lists compatible with a Lynx LM1800 liquid handler in addition to custom scripts automating cell passaging and sampling. This approach yielded an end-to-end 36 day CLD workflow capable of generating high-titer cell lines for multiple complex antibody structures. Here, we open-access our trained models, user interface, and Lynx automation scripts to provide a modular toolkit useful for clonal cell line engineering projects. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=153 SRC="FIGDIR/small/703387v1_ufig1.gif" ALT="Figure 1"> View larger version (51K): org.highwire.dtl.DTLVardef@1f72e70org.highwire.dtl.DTLVardef@109c54dorg.highwire.dtl.DTLVardef@7867b1org.highwire.dtl.DTLVardef@dfa61e_HPS_FORMAT_FIGEXP M_FIG C_FIG

cell biology↗

An Efficient Workflow for CHO Cell Genome Engineering with OpenCRISPR-1

The effective titer and quality of biopharmaceutical products can be enhanced by genome engineering of producer cell lines; however, licensing constraints often limit nuclease utility. Here, we validate OpenCRISPR-1 in CHO cells by achieving [≥]70% INDEL efficiency across multiple genes. We demonstrate biallelic Fut8 knockout in monoclonal cell lines with 31% efficiency and quadruplex knockout of lipases at 7% efficiency using a 39-day workflow. This work highlights the potential applications for democratized nucleases in host cell engineering.

bioengineering↗

CD388: A universally protective Drug-Fc Conjugate that targets influenza virus neuraminidase

The ability of the influenza virus to elude humoral immunity by rapid antigenic shift presents a sustained and urgent threat to human health. Globally, seasonal influenza causes an estimated 3 - 5 million cases of severe disease and 300,000 - 500,000 deaths annually, with increased potential for mortality during pandemics1. The recent outbreaks of avian H5N1 in bird populations, subsequent spread to cattle and appearance in humans, highlight the need for effective broad-spectrum influenza antivirals for treatment, prophylaxis and pandemic preparedness. The narrow, strain-specific immunity induced by current seasonal vaccines and mismatches between vaccine strains and circulating viruses result in limited vaccine effectiveness (VE) rates2. Furthermore, VE is even lower in immune-compromised and - senescent populations3. Strategies that provide durable, universal influenza protection in healthy and high-risk populations are urgently needed. Here, we describe CD388, a first-in-class antiviral drug-Fc conjugate (DFC) in clinical trials for seasonal influenza prevention (NCT05285137 and NCT05523089). CD388 comprises a multivalent small molecule inhibitor of influenza virus neuraminidase (NA) linked to a CH1-Fc hybrid domain of human IgG1 engineered for extended half-life. CD388 demonstrated potent, universal activity across influenza A and B viruses, including high pathogenicity and NA resistant strains, a low potential for resistance development, and efficacy in lethal mouse infection models. CD388 is the first therapeutic with the potential for universal prevention of influenza A and B in healthy and high-risk populations.

microbiology↗