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Scalable High Throughput Selection From Phage-displayed Synthetic Antibody Libraries
Published on: January 17, 2015
Programmable nanobody circuits for cell selection
Nathan B Wang1, Albert Blanch-Asensio1, Hannah Cevasco2
1Department of Chemical Engineering, MIT, Cambridge, MA 02139, USA.
Biorxiv : the Preprint Server for Biology
|May 13, 2026
Summary
We developed a novel protein-based circuit, the Destabilized-nanobody Antigen Selection and Identification Tool (DASIT), for efficient, antigen-specific cell selection. DASIT enables scalable, FACS-free cell enrichment for diverse applications in genome engineering and cell therapy manufacturing.
Area of Science:
- Biotechnology
- Molecular Biology
- Cell Biology
Background:
- Efficient isolation of specific cell populations is crucial for advancing genome engineering, pooled screening, and cell therapy manufacturing.
- Current methods often face bottlenecks in scalability and specificity, hindering progress in these fields.
Purpose of the Study:
- To develop a novel protein-based circuit for antigen-specific cell selection, named DASIT (Destabilized-nanobody Antigen Selection and Identification Tool).
- To demonstrate DASIT's capability for scalable, FACS-free cell enrichment across various cell types and applications.
- To establish an automation-compatible architecture for complex genetic engineering and cell manufacturing processes.
Main Methods:
- Engineered a destabilized nanobody fused to an antibiotic resistance protein, forming the DASIT circuit.
- Utilized antigen binding to stabilize the DASIT circuit, creating a selectable signal for antigen-positive cells.
- Applied DASIT for multiplexed, logic-gated integration in human iPSCs, high-throughput CRISPR screening, and phenotypic selection of engineered neurons.
Main Results:
- DASIT enables robust selection of antigen-expressing cells across different cell types and antigens.
- The system supports both stable integration and transient delivery, allowing recyclable selection without permanent genomic modification.
- Demonstrated scalable, FACS-free enrichment in challenging applications, including iPSC engineering, CRISPR screening, and neuronal cell therapy.
Conclusions:
- DASIT provides an efficient and scalable solution for antigen-specific cell selection, overcoming key bottlenecks in cell engineering and manufacturing.
- Decoupling selection from vector integration offers a flexible and automation-compatible platform for multistep genome engineering and large-scale cell production.
- DASIT facilitates advancements in high-throughput screening, cell therapy, and precision genome engineering.
