Cell Transient Transfection Expression Service

Transient transfection of mammalian cells expresses foreign DNA for a limited period without integrating the DNA into the genome and replicating a new gene. As the most commonly used strategy for mammalian protein expression, the transient transfection service provided by Biologics International Corp. (BIC) produces 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.

Key Features

  • High transfection efficiency: High transfection efficiency is achieved through the combination of electroporation transfection and chemical transfection.
  • Gene optimization technology: Our self-designed MaxCodonTM software can optimize codon mRNA secondary structure and ribosome binding sites to improve protein expression efficiency.
  • Cells with long life: The optimized transfection method can effectively prolong the life cycle of the host cells, increasing the accumulation time of target proteins.

Transient Protein and Antibody Services

Transient Transfection Process

 

Service process 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

Cell Transient Transfection Flow Chart:

Transient Transfection Service Procedure

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
(B) mRNA with optimized structure
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

Case Study

Comparison of the expression levels before and after optimization.

transient-case

Figure on the left clearly shows a significant increase in protein expression with optimized vector and cells.

Contact Us

phone+1 (317) 703-0614
fax +1 (855) 427-1516
 For questions and/or a price quote, please inquire online or fill in a pre-order form.
contact

captcha