Phage display antibody library technology is often difficult to meet the needs of antibody-drug development. In recent years, the mammalian cell surface display system has become an important direction for the development of human antibodies. The process of protein folding, secretion and post-translational modification in mammalian cells is the closest to that in the human body.
The application of the mammalian cell surface display system can display full-length human antibodies, which has the potential to exert the effect function of antibody constant region. The traditional full-length antibody library construction technology is not mature, and the construction efficiency is low. At Creative BioMart, our improved technology platform can complete the construction of 1010 full-length antibody library in 2 weeks.
| Convectional mammalian antibody library | Creative BioMart's full-length antibody library | |
| Construction efficiency | low | high |
| Library affinity | low | high |
| Library specificity | low | high |
| Maximum library size | 106 | 1.4 x 1010 |
| Time consumption | More than 3 weeks | 2 weeks |
Isolation of human antibodies by mammalian cell display (Beerli, 2008)
DH5α Calcium sensing cells and CHO cells are involved in the construction of antibody library by default. If you have other requirements, please specify them before the project starts. We will design three sets of heavy chain primers and three sets of light chain primers according to your target fragment. Rapid amplification can be achieved by six PCR reactions.
After receiving your samples or sequences submitted online, we will process and evaluate the sequences, and synthesize and design primers. The procedure of antibody library construction includes
We have constructed a large-scale expression vector. CMV promoter, PDGFR transmembrane sequence and full-length IgGI heavy chain gene of human antibody were included in the vector. There were ampicillin screening markers and multiple restriction sites on the vector.
To design and apply a set of primers with good amplification efficiency is the key to improving the library capacity and maintaining the diversity of antibody library. Three sets of primers will be used to amplify the whole set of the variable region of heavy chain and full-length kappa light chain respectively. After purification and recovery, they are mixed in equal proportion to balance the proportion of each subtype DNA in the constructed antibody gene library, so as to ensure the diversity of antibody library. We will randomly select clones from the antibody gene library for sequencing to ensure that the sequence accuracy is more than 90% and there is no repetitive sequence.
Customers who need our assistance are more than welcome to contact us or submit an online inquiry. Our professional team will provide you with detailed guidance and solution ASAP.
Reference
1. Beerli RR, Bauer M, Buser RB, Gwerder M, Muntwiler S, Maurer P, Saudan P, Bachmann MF. Isolation of human monoclonal antibodies by mammalian cell display. Proc Natl Acad Sci USA. 2008 Sep 23;105(38):14336-41.
