Vector Retrofits

Vector Retrofits

Highly expressed • Durable • Antibiotic free • Proven

NTC’s antibiotic free Nanoplasmids™ are easily substituted for conventional plasmids, minicircles, doggy bones, etc., for use with all major therapeutic vector groups. The Nanoplasmid™ backbone has profound and unexpected beneficial effects on performance (expression levels and duration). The Nanoplasmid™ is now available for use in: Lenti vectors, CAR-T cell vectors, AAV, transposons, etc. NTC’s RNA-OUT sucrose selection is a regulatory agency preferred antibiotic-free backbone, and has already been approved and used successfully in a number of first-in-human clinical trials.

Proud to offer retrofit services, including verification and DNA production.

Send us 10 micrograms of your plasmid (on filter paper or in liquid form) and NTC will swap your current plasmid backbone for the Nanoplasmid one, verify the construct, and return the desired quantity of the amplified product, along with the R6K host strain. Its small size also increases potency and reduces the cost of producing plasmid copy number equivalents
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Example

Retrofitting a pVAX plasmid (1) with RNA-OUT (antibiotic free) selectable marker (2), or (3) with Nanoplasmid™ (RNA-OUT plus R6K ori), or (4)with Nanoplasmid™ plus BGH poly(A), dramatically improves expression, up to ten (or more) fold. Scale: Fluorescence Units (FU).

pVAX Nanoplasmid™ Retrofits

 

No. Vector Intron ORI-Marker A549 EGFP-FU +/- SD HEK EGFP-FU +/- SD
1 pVAX-EGFP-Kan (-) pUC-Kan 658 68 3797 351
2 pVAX-EGFP-RNA-OUT (-) pUC-RNA-OUT 1737 79 15974 934
3 pVAX-EGFP-Nanoplsamid™ (-) R6K-RNA-OUT 3142 242 24155 1051
4 NTC93835R-EGFP-Nanoplsamid™ (-) R6K-RNA-OUT 4379 389 38421 2528

In summary, the Nanoplasmid (RNA-OUT selectable marker plus R6K origin) replaces antibiotics with sucrose selection, and replaces the promiscuous pUC origin with the host-restricted R6K one. It is:

  • Regulatory compliant

  • Smaller in size (452bp) than conventional marker/ORIs, therefore increasing potency and decreasing material cost

  • Easily swappable into most types of viral and nonviral vectors

  • HyperGRO compatible for high yield Nanoplasmid manufacturing
  • Purified using conventional plasmid DNA downstream purification processes, and does not require post-production modification (such as minicircles), and is therefore the ideal vector for products that are destined for clinical use

  • Dramatically improves expression levels and duration of expression compared to Kanamycin vectors

Nanoplasmid™ is an ideal vector: Used by over 50 gene therapy companies worldwide

Small Backbone • Fast • Economical • Antibiotic-Free • Scalable