• Plasmid Fermentation E. coli Cell Paste, for highest yield plasmid DNA using the HyperGRO™ process: $3,600/10L.

    DNA Fermentation Process

    Do you want to purify pDNA with your own downstream process, but you also want to take advantage of NTC’s HyperGRO™ fed-batch fermentation technology? If so, you may want to order your plasmid as frozen, concentrated cell paste, directly from NTC. NTC will perform a 10L fed-batch HyperGRO™ fermentation using its optimal process (typical yields: 300mg-2.6g/L), and send you all of the paste from a 10L fermentation, for $3,600.

    HyperGRO™ - E. coli Cell Paste

    E. coli cell paste, Plasmid containing cell paste can be frozen and used at any time to isolate HyperGRO plasmid DNA.

     

    NTC offers full downstream purification services

    Other Options:

    • Large scale plasmid DNA Production

    • Small Scale Plasmid DNA

    Site licenses: For contract research facilities or for in-house production.

    Biotech Suppliers:

    Consider out-sourcing your company's entire plasmid production effort to NTC, saving you lots of money and allowing you to re-prioritize your staff. Your orders will receive creful management from our knowledgeable and experienced professionals. Timely delivery and high quality products are guaranteed with NTC' industrial outsource program. Perfect for biotech suppliers and vector banks.

    Process Development Fermentations:

    HyperGRO plasmid DNA fermentation

    Paste fermentations are thus incorporated into process development, and may be ordered for that purpose alone, or they may be combined with purification in order to obtain pDNA.  If you choose to have NTC purify the DNA, the cost of the fermentation ($3,600) will be deducted from the regular cost of purified DNA.

    Learn more about NTC's HyperGRO™ Plasmid DNA Manufacturing Platform (click here)

    HyperGRO™ Publications

    Plasmid DNA Fermentation Strain and Process-Specific Effects on Vector Yield, Quality and Transgene Expression, Carnes, A.E., Luke, J.M., Vincent, J.M., Schukar, A., Anderson, S., Hodgson, C.P., Williams, J.A. Biotechnol. Bioeng. (2011). Feb;108(2):354-63.


    Plasmid DNA Production Combining Antibiotic-free Selection, Inducible High Yield Fermentation, and Novel Autolytic Purification, Carnes, AE, Hodgson, CP, Luke, JM, Vincent, JM, Williams, JA, Biotechnol. Bioeng. 104:505-515 (2009)


    Generic plasmid DNA Production Platform Incorporating Low Metabolic Burden Seed-stock and Fed-batch Fermentation Processes, James A Williams *, Jeremy Luke, Sarah Langtry, Sheryl Anderson, Clague P Hodgson, Aaron E Carnes, Nature Technology Corporation, Lincoln, NE, USA. Biotechnol. Bioeng. (2009) 103:1129-114

     

    Rapid process development for high yielding plasmid DNA fed-batch fermentation. Williams, J.A., Hodgson, C.P., and Carnes, A.E. (2009).BioPharm International 22(11):46-51

     

    Inducible Escherichia coli fermentation for increased plasmid DNA production, Carnes AE, Hodgson CP, Williams, JA. (2006) Biotechnol. Appl. Biochem. 45:155-166.

  • DNA Manufacturing

    HyperGRO fed batch plasmid DNA production

    Advantages of NTC’s Plasmid Manufacturing Capabilities

    • Highest yields ( > 2.6 g/L)

    • Improved % supercoiled monomer using reduced metabolic burden process

    • Highest quality purification (>95% supercoiled plasmid DNA)

    • Highest plasmid yields per unit biomass

    • Lowest cost per unit DNA

    • Vector to process matching for highest overall performance

    • Improved yield of toxic or unstable plasmids

    Save 20% of large scale production cost with NTC’s high-yield vectors

     

    Production of highly-purified research-grade plasmid DNA at NTC’s Lincoln, Nebraska, USA plant range from 1mg to multi-gram scale, and production capabilities for GMP-grade DNA at NTC’s CMO partners range from a hundred milligrams upward.

    Small to large scale plasmid DNA production

    NTC’s In-house pDNA services

    • Small- and large-scale production and purification of research-grade pDNA (1mg-multi-gram)

    • Quality testing of pDNA products (endotoxin [LAL]; residual RNA [gel], DNA [qPCR], protein [modified BCA testing])

    • Production of plasmid-enriched E. coli cell paste for downstream processing in client facilities

    E. coli strain engineering, for production hosting of specific toxic plasmids

    • Plasmid backbone engineering for substitution of antibiotic-free (markerless) or high copy plasmid backbones

    • Low metabolic burden technology for stable production of toxic or unstable plasmids

    • Production of Research Cell Banks (RCB) using low metabolic burden technology

    • Transfer of RCB to GMP facility for certification as Master and Working Cell Banks

    • Technology Transfer of HyperGRO™ process to CMOs and end-user facilities for best-in-class performance

  • Small Scale Plasmid Production

    Small Scale Plasmid Production

    DNA Manufacturing,  mg scale plasmid DNA prep services

    Small Scale Certificate of Analysis

    • Spectrophotometric analysis (A260/280)

    • Quantity

    • Concentration (usually 0.5mg/ml, in TE buffer)

    • Volume

    • Gel photo (digested and undigested DNA, ladder)

    • Qualitative analysis (pass/fail)

    NTC is an industry leading provider of highly-purified plasmids. Ordering highest quality plasmid DNA from NTC is a fast, easy and economical process.

     

    Timeline: First time orders require 2-3 weeks. Reorders can be from 1 day to two weeks, depending upon the arrangements made (just in time re-ordering is by prior agreement).

    NTC will:

    • Transform the production host.

    • Prepare an archival glycerol stock using "low metabolic burden technology" ,

    • Grow, and purify the desired quantity of pDNA, using endotoxin-free reagents and procedures.

    • Formulate the DNA according to specification (default is 0.5mg/ml in TE buffer).

    • A small-scale certificate of analysis will be provided, including: gel photo (digested and undigested DNA, ladder), concentration and quantity.

    DNA Manufactring: certificate of analysis for a small scale plasmid DNA preparation.

    Small Scale C of A

    In addition, NTC will retain an archival lot sample, and a glycerol stock of your plasmid for future reorders (at least one year retention).

  • Large Scale Plasmid Production

    Large Scale Plasmid Production

    DNA Manufactring, NTC gram scale plasmid DNA prep services

    At no additional charge, Each

    endotoxin-free DNA prep includes:

    1) Transformation


    2) Glycerol stock archiving


    3) Lot sample archive


    4) Custom formulation


    5) Certificate of Analysis (C of A)

    In addition, NTC will retain an archival lot sample, and a glycerol stock of your plasmid for future reorders (at least one year retention).

    Large scale DNA plasmids at NTC are grown in a fed-batch fermentation process, permitting control over feed rate, pH, temperature, and dissolved oxygen. NTC’s patented HyperGRO™ fermentation process and media has produced record pDNA yields of >2.6g/L, and typically produce 0.5-1.5g/L using a high copy pUC, or similar origin, and a recA-, relA-, endA- host strain, such as DH5 alpha. Highly concentrated HyperGRO™ plasmid preparations naturally result in cleaner, more highly-purified DNA (due to dramatically reduced biomass), and more cost-effective production as well. (See related molecular services below)

     

    Large Scale Plasmid DNA Purification and Production (100mg-multi gram) Low Endotoxin Plasmid DNA Purification and Processing

     

    Also:

    A large-scale certificate of analysis will be provided including gel photo (digested and undigested DNA, ladder), concentration and quantity:

    Large Scale Certificate of Analysis, (pdf) including:

    ·  Spectrophotometric analysis (A260/280)

    ·  Quantity

    ·  Concentration (usually 0.5mg/ml, in TE buffer)

    ·  Volume

    ·  Gel photo

    ·  Residual RNA (gel assay)

    ·  Residual genomic E. coli DNA (qPCR)

    DNA Manufactring, NTC large scale plasmid DNA certificate of analysis.

    Large Scale C of A

  • Additional Testing

    Additional Testing

    DNA Manufacturing: Additional Testing
  • Cost Comparison

    Cost Comparison

    DNA Manufactring, cost comparison
  • Ordering

    Ordering Plasmid DNA from NTC

    NTC is an industry leading provider of highly-purified plasmids. Ordering plasmids from NTC is a fast and easy 2 step process, and there is no where else you will receive the attention to detail and great service for the cost.

     

    Timeline: First time orders require 2-3 weeks. Reorders can be from 1 day to two weeks, depending upon the arrangements made (just in time re-ordering is by prior agreement).

     

    1) Call NTC for a quotation, (402) 323-6289. Best pricing is available based on quantity ordered and copy number. Most high-copy plasmids have a pUC origin of replication, or equivalent, and result in the lowest cost. NTC is a longtime industry leader in quality and cost-effectiveness.

     

    2) Send NTC 1microgram of your plasmid DNA, spotted on Whatman 3MM or FTE filter paper, together with NTC's Plasmid DNA Order Form filled out. (The form is designed to prompt you for all information needed for propagation and testing.)

     

    NTC will:

    • Transform the production host.

    • Prepare a glycerol stock using "low metabolic burden technology"

    • Grow, and purify the desired quantity of pDNA, using endotoxin-free reagents and procedures.

    • At no additional cost, NTC will also formulate the DNA according to specification (default is 0.5mg/ml in TE buffer).

    • A large-scale certificate of analysis will be provided, including: gel photo (digested and undigested DNA, ladder); concentration and quantity; information and testing for residual: RNA, genomic DNA (qPCR). Upon request, additional testing for protein, endotoxin [LAL assay] and bioburden are available. (see Additional Testing below)

  • Linear Vectors

    Linear Vectors

    Custom linearized vectors for high-throughput genomics and screening

    Effectively solving the inefficiencies of gel isolation (ethidium and UV damage) and background (reduced 4-5 logs).

     

    The initial and limiting factor in a library or high-throughput cloning project is the vector.  Careful preparation of the vector using a proprietory process gives the highest cloning efficiency, and the lowest background (parental and empty clones).

    Linear Vectors, NTC uses ethidium bromide free and UV free methods of DNA isolation from gels, thus retaining highest transfection efficiency.

    Unlike over-digestion, which does not rid the preparation of unwanted, intact vector, NTC's process results in clean, low-background linearized vectors that you can use with confidence, knowing that the product wil give you the highest cloning efficiency possible.

  • DNA Fermentation Process

    DNA Fermentation Process

    NTC’s High Yield HyperGRO™ plasmid DNA fermentation process is ideal for making high-purity DNA vaccines and gene therapeutics.

    Comparison of shake flask vs. fed batch fermentation plasmid yields
    HyperGRO fed batch fermentation

    Fig. B) HyperGRO™ fermentation process schematic.

    DNA fermentation: HyperGRO DNA fermentation yield

    Fig. A) Yield comparison, typical vs. HyperGRO™.

    Fig. C) Fed-batch HyperGRO™ fermentation using NTC7264 backbone with 30-42C temperature shift, yield 1,500mg/L (left scale, mg/L

    NTC customers have several options:

     

    1) Complete Plasmid DNA Purification Services: Small scale (1-99mg), and Large scale (100mg-kg) custom-purified, research-grade DNA is available directly from NTC. Orders 500mg and up are generally made using HyperGRO™. In addition, GMP grade DNA is available through NTC’s GMP manufacturing partners (inquire).

     

    2) Process Development/Cell Paste: In order to see if your plasmid is compatible with the HyperGRO™ fed-batch fermentation process, shake-down runs are available, in which an NTC engineer will optimize fed-batch fermentation for your host cell/plasmid combinations. We will concentrate, freeze and provide you with the cell paste from optimal runs if you would like to purify it elsewhere, or NTC can complete the purification.

     

    3) Technology Transfer to your facility: An NTC engineer will set up the process in your facility and train technical staff to make the world’s best DNA. NTC will provide all technic

    al documentation.

VECTORS & CELL LINES

Nanoplasmids™

RNA-OUT Retrofit
Mammalian Expression Vectors

Antibiotic Free Vectors

DNA Vaccines

Rig-I Activating Vectors

Recombineering Vectors

pVEX™- E. coli Expression Vector

Anti-Silencing Elements

Gene Silencing

Transient

Retroviral, stable

Cell Lines

Autolytic E. coli Cell Lines

dcm- Strains

 

PLASMID MANUFACTURING

HyperGRO™, E. coli Cell paste

Contract Plasmid Manufacturing

Small Scale Large Scale

Additional Testing

Order Form

Linear Vectors

 

CLONING SERVICES & QC

Rapid VACC™ Precision Cloning

Plasmid Detox Services

Contract QC

Vector Retrofit Services

Swap Marker or Promoter

E. coli Genomic DNA Testing

Residual RNA qPCR

Residual RNA gel

Supercoil Assay

Lymulus Endotoxin Testing

 

PROTEIN PRODUCTS

Thermostable Protein Expression

HN-Flagellin Protein

HN-TST-HA2 Protein

HN-TST-Flagellin Protein

HYPERTAQ™ DNA Polymerase

Custom Recombinant Proteins

Fluorescent Proteins

TECHNOLOGY & CONSULTING

Process Development

Plasmid Fermentation & Purification

Protein Fermentation & Purification

High yield plasmid Fermentation

ABOUT NTC

Patents

Publications

Management Team

Board

Investor Information