VectorGPS® Expression Optimization Technology

Discover the vector that works best in your expression system. Vectorology™ efficiently explores vector element determinants of protein expression.

Create the vector that works best in your expression system. VectorGPS uses Design-of-Experiment (DoE) algorithms to build testable numbers of vectors from sets of control elements, Vectorology™ and advanced machine learning to assess the contribution of each element to vector performance. Critical vector elements perform differently in different systems, so instead of using vectors packed with fossil sequences put there by graduate students twenty years ago for reasons no one remembers, let ATUM help you create the custom vector you need.


Recommended Vector Selection Process:


Expression Vector
1. Single Expression Vector
Already know exactly which combination of elements you want? Use the Vector Selector on the order tab to choose one of our ready-to-use vectors.


Vector Panel Icon
2. Expression Panel
Not sure how your protein will behave? Test your gene in several vectors with a variety of properties most likely to increase expression. ATUM offers pre-selected panels for Bacterial Expression, Bacterial Secretion or Mammalian Expression (Transient and Stable), or you can create your own to explore the properties you think are most important.


VectorGPS Icon
3. VectorGPS
Want the very best vector for your system? Working in a non-standard expression system, a particularly difficult protein, or need even higher yields of active protein? ATUM will create unique combinations and configurations of vector components using our machine learning technology, to create a custom vector exactly suited to your needs.

Two Ways to Order:

1. Catalog Expression Panels (Ready-to-Use)

Test your gene in several ready-to-use vectors with a variety of properties most likely to increase expression. ATUM offers pre-selected panels for Bacterial Expression, Bacterial Secretion or Mammalian Expression (Transient and Stable).

2. Custom VectorGPS Project

Let ATUM do the work for you.
We will apply our Vectorology and Bioengineering GPS to design a custom vector tailored for your expression system. Combinations of vector components are designed so that elements are varied systematically and independently. We then use advanced machine learning algorithms to assess the contribution of each element to vector performance for your protein.


Catalog Expression Panels (Ready-to-Use):

ATUM has pre-selected panels of vectors to facilitate optimization of vector components for bacterial expression, bacterial secretion, transient mammalian expression, and stable mammalian expression. You can also build your own custom panel by selecting any set of vectors. The quickest and easiest way to start vector optimization is to clone your gene into a Panel set of Electra System vectors, or you can request your gene cloned into a panel of vectors.

Vector Selector


Custom VectorGPS Project:

Let ATUM do the work for you.
Contact ATUM, and we will apply our Vectorology and Bioengineering GPS technology to design a custom vector tailored for your expression system. Combinations of vector components are designed so that elements are varied systematically and independently. We then use advanced machine learning algorithms to assess the contribution of each element to vector performance for your protein.

VectorGPS for Increased Expression in Bacteria

Variations in promoter, ribosome binding site, N- or C-terminal fusions, origin of replication and even selectable marker can have pronounced impacts on protein expression in E. coli.

VectorGPS Bacterial Elements Diagram

 

In addition to the opportunity to improve expression vectors for E. coli, the DoE, Vectorology, and machine learning of our GPS technology have enormous potential for optimizing expression for other, less well studied, bacterial systems. Contact info@atum.bio to discuss a VectorGPS project for your system.


Protein Expression from Different Promoters


Let us test the Panel for you: ATUM Expression Test
Different promoters work better for different proteins. Expression of three proteins, phi29 (~61 kD), cutinase (~22 kD) and DasherGFP (~26 kD), under control of different promoters, T5 and T7 (IPTG-inducible), rham (rhamnose inducible )and phoA (inducible by phosphate starvation), in low and high copy vectors is shown. Expression values shown on the y-axis are measurements of expressed protein band densities from a SDS-PAGE gel using BSA as standard.
*DasherGFP fluorescence measured is lower with T7 and rham expressed GFP indicating that a fraction of total protein is incompletely folded protein.

Effectiveness of Solubility Tags is Protein Dependent

Expression of total and soluble protein obtained using different tags. Three proteins encoded by genes A (~48 kD), B (~49 kD) and C (~59 kD), having shown poor solubility, were expressed in IPTG-inducible T5 promoter E. coli expression vectors (pD441-XXX) with various N-terminal solubility tags: MBP, GST, PpiB, and Fh8; 6XHis was included as a negative control. Cultures were grown at room temperature (RT) and 37°C. Overall, higher soluble protein expression was observed at RT (data shown). Percent solubility is shown along the Y-axis and the size of the circles corresponds to protein yield shown in nmole/µl. Legend shows approximate yield of protein based on circle size.

Mammalian vector systems in particular, have a large number of vector elements that contribute to vector performance. Viral vectors such as lentiviral and adeno-associated viral (AAV) vectors can suffer from additional constraints of sequence and size. We have identified vector elements in bacterial, mammalian, lentiviral and adeno-associated viral systems that show enhanced expression and/or integration properties.

VectorGPS vector map


Featured in:


Example: Application to Mammalian Transfection Vectors:

Using ATUM Vectorology, forty-eight vectors were constructed featuring combinations of the following sequence elements. The vectors were used to express DasherGFP in HEK293 cells and CHO cells. Each vector was transfected in triplicate, expression was measured by harvesting cells 72 hours after transfection, lysing them and measuring fluorescence in a fluorimeter.

Enhancer Promoter Intron PolyA Amplification
CMV Chick actin Chick actin Β globin EBNA
EF1α CMV CMV A BGH OriP
SV40 EF1α CMV C HSV-TK SV40 T
Synthetic GAPDH EF1α SV40 early  
  HSV-TK Synthetic SV40 late  
  HTLV   Synthetic  
  MC1      
  PGK      
  SV40      
  Ubiquitin B      

 

Because the vector components were varied systematically, we were able to use machine learning to assess the contribution of each element to expression in HEK and CHO. Most promoters and polyA sequences performed similarly in the two systems. The effect of viral amplification sequences was dramatically different: positive or neutral in HEK and strongly negative in CHO.

 

CHO HEK Expression Vector Variables

Systematic sampling of mammalian vector elements. Machine learning algorithms identified vector components that contributed to expression of DasherGFP in HEK293 (red bars) or CHO (blue bars) cells. Positive contributions have positive regression weights, the magnitude of the weight corresponds to the magnitude of the effect of the element.

A pre-built set of vectors is an economical place to begin a VectorGPS study. It gives a rapid overview of the variability of expression in your system that can result from different combinations of common vector components.


Transient Vector Performance Varies with Cell Type

DasherGFP Expression in Transiently Transfected HEK293 and CHO Cells



Mammalian Transient Vector Discount
BuyGet
2 10% off
3 20% off
4 or more 30% off

 

Legend

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DasherGFP expression in transiently transfected cells. HEK293 (adherent), Expi293™ (suspension), CHO-K1 (adherent), Freestyle™ CHO-S (suspension), and ExpiCHO™ (suspension) cells were transfected at 70-80% confluency using Lipofectamine 2000; transfections were carried out in triplicate and incubated for 72 hours post-transfection. Cells were lysed using M-PER and expression measured on a fluorimeter. Yellow data points are shown for comparison.

Stable Vectors Also Show Good Transient Expression

DasherGFP Expression in Stably Transfected CHO Cells, Transient Expression in HEK293

Mammalian Stable Vector Discount
BuyGet
2 10% off
3 20% off
4 or more 30% off

 

Legend

Available (mouse over to see options) In my cart/favorites Email info@atum.bio formore information Labeled for comparison Error bar
DasherGFP expression in stably transfected CHO cells and transient expression in HEK293 cells. Cells were transfected at 70-80% confluency using Lipofectamine 2000; transfections were carried out in triplicate in HEK293 cells and CHO cells. Selection media with 5 µg/ml Puromycin was added 72 hours post-transfection, grown for an additional 72 hours post-selection and media changed to complete growth media for a total of 14 days (2 passages in complete growth media) for CHO cells. DasherGFP transient expression was measured 72 hours post-transfection in HEK293 cells. Cells were lysed using M-PER and expression measured on a fluorometer.

Consult with a VectorGPS Specialist Today

To determine the best possible option for your research project please contact our VectorGPS Specialists at +1 877 362 8646 or info@atum.bio.