Cell Culture and Transient Transfection

Creative Bioarray can use appropriate transfection methods to meet customers' transient transfection needs. We have established a mature agreement to provide instant transfection technical services with fast turnaround time according to your needs.

Cell Culture in Transient Transfection

The transient expression means that the expression vector of the cloned foreign gene can express the foreign protein without being integrated into the host cell genome, and the recombinant protein can be quickly prepared in a short time. Transient expression technology can be used for the research of gene function using RNA interference technology and the development of recombinant monoclonal antibodies and recombinant protein drugs. In transient transfection, the introduced nucleic acid only exists in the cell for a limited time and does not integrate into the genome. Therefore, transiently transfected genetic material will not be passed on from generation to generation during cell division and may be lost due to environmental factors or diluted during cell division. However, the transfected genetic material requires the use of biochemical, biological or biophysical transfection methods for nucleic acid transfection, and it is transient (expression time is limited).

Fig 1. Flow chart showing the entire process from thaw to transfection. (Arena T A, et al. 2018)

Cell Culture and Transient Transfection Services

Transient gene expression (TGE) is a rapid method for the production of recombinant proteins in mammalian cells. Our transient transfection and cell culture services are mainly for the situation where the vector has been constructed. Customers only need to provide the constructed vector and select the expression host cell or choose our vector construction service. This service provides from transfection to subsequent 2-4 weeks of culture. Our transient transfection service platform provides services such as empty cell culture, transient transfection and post-transfection culture according to the platform technology. The specific service process includes cell resuscitation, transfection, transfection detection and cell counting. This service can provide transient transfection reports, post-transfection culture processes and transfection culture supernatant or purified product.

Fig 2. Our transient transfection and cell culture service process.

Transfection Method and Quality Control

• Transfection method
• Different transfection methods have different characteristics. We determine the best transfection method according to the specific needs of customers and experimental limitations during transfection experiments.
• Commonly used cell transfection methods include electroporation, microinjection, gene gun, liposome transfection, calcium phosphate co-precipitation method, protoplast transfection, and virus-mediated transfection.
• Precautions
• Cell state. Make the cells in a good growth state before transfection.
• Transfection reagent. Different transfection reagents are used for different purposes/fragment types, and our platform contains a variety of transfection reagent options.
• Plasmid purity (A260/A280 ratio (1.7-1.9)) and plasmid concentration (the plasmid concentration for cell transfection is not less than 500ng/ul, and the plasmid concentration for virus packaging is not less than 0.8ug/ul).
• Suitable endotoxin removal.
• We will adjust the appropriate transfection conditions according to different projects, and analyze the specific conditions to ensure the quality of service.

Fig 3. The quality control part of our transient transfection service.

Creative Bioarray provides transient transfection and cell culture technical services for researchers who have constructed vectors, helping our customers to quickly establish good expression. You will benefit from our technical expertise and state-of-the-art facilities. Our scientific team will work with you to find the best solution for rapid transient gene expression.

References:

1. Arena T A, Harms P D, Wong A W. High throughput transfection of hek293 cells for transient protein production[M]//Recombinant Protein Expression in Mammalian Cells. Humana Press, New York, NY, 2018: 179-187.
2. Geisse S, Voedisch B. Transient expression technologies: past, present, and future[J]. Therapeutic proteins, 2012: 203-219.
3. Büssow K. Stable mammalian producer cell lines for structural biology[J]. Current opinion in structural biology, 2015, 32: 81-90.