Transient Transfection Service

Transient transfection of mammalian cells expresses foreign DNA for a limited period time without integrating it into their genome and replicating new gene. As the most commonly used strategy for mammalian protein expression, transient transfection service provided by BiologicsCorp produce microgram quantities of high-quality proteins rapidly. The strategy has been applied to short-term transient expression of genes and small-scale protein production for research and development purpose.

Transient Transfection Service Key features

  • High yield: capable of producing >200mg/liter in our transient transfection system
  • High transfection efficiency: multiple optimizations to significantly improve transient transfection efficiency compared with regular process
  • Competitive price: start from $2,900

 

Transient Transfection Process

Transient Transfection Service Procedure

Cat. PS1314 Procedures Price Deliverables Timeline
Gene synthesis
  • Codon optimization
  • Gene synthesis
  • Subcloning & Max-Prep
Please Inquire Cloning vector 3 weeks
Transient expression and purification
  • Expression detection
  • 1L pilot-scale expression
  • Purification
  • QC by SDS-PAGE and WB
Please Inquire
  • Purified recombinant antibody
  • QC data and final report
4~6 weeks

 

Transfection Expression Level Improvement

mRNA structure and codon optimization

Increasing stability of the transgene mRNA was proved to be key factors for high quality transient mammalian cell transfection. Stem-loop structure may encode features that cause mRNAs to be labile, which severely impact process of gene transcription and translation.

Our mRNA structure and codon optimization tools refer the same amino acid to multiple triplet codons. It also opens stem-loop structures in mRNA sequence and subsequently increase transient expression yield.

(A) mRNA with stem-loop structure
mRNA with stem-loop structure for transient expression
(B) mRNA with optimized structure
mRNA with optimized structure for transient expression
Figure 2: Comparison of mRNA sequences with stem-loop and optimized structure

After transient expression, the optimized constructs achieved significantly increased high level expression compared with a previously wild type product.

(A) Wild type protein(B) Optimized protein

Comparison of wildtype protein expression and optimized protein expressions

Figure 3: Transient expression comparisons of wild type protein and optimized protein

 

Transient Transfection Efficiency Improvement

    • Special transfection reagent

Our in-house transfection reagent is designed to improve post transfection efficiency. It is formulated for transient transfection in mammalian expression system and generally presents significant advanced transfection efficiency than regular reagent.

    • Electroporation transfection

In recent years, electroporation has become the preferred choice due to its high efficiency. The technique is a procedure gaining in popularity for gene transfer using high-voltage electric shocks to introduce DNA into cells. The DNA attached to feeder layer cells quickly and formed fast-growing colonies. The simple and efficient method yields a high frequency of protein transient expression.

(A)ControlConventional method

Control A
Conventional method

(B)ControlOptimized strategy I

Control B
Optimized strategy 1

(C)ControlOptimized strategy II

Control C
Optimized strategy 2
Fig. 4: Compared with conventional method, our technology achieves higher transient transfection efficiency.
  • Conventional method: lipid-based transient transfection
  • Optimized strategy I
    • Special transfection reagent
    • Electroporation-mediated transient expression
  • Optimized strategy II
    • Optimized plasmid
    • Special transfection reagent
    • Electroporation-mediated transient expression

Contact us

  +1 (317) 703-0614
  +1 (855) 427-1516

* To get a price quotation, please fill the protein service pre-order form and email to Sales@BiologicsCorp.com.