Creative Biolabs has combined the high-efficient DNA immunization strategies with our advanced phage display technology to generate high specific antibodies for the target of interest. Our scientists have extensive knowledge and experience in immune library construction, screening, and antibody manufacturing.
Antibody Phage Display
Phage or bacteriophage refers to viruses that infect and amplify in bacteria cells. Phage display technique was first established by Smith, GP in 1985 and further developed by a number of researchers. By genetically packaging encoding DNA into phagemid vector, antibody/protein can be fused to bacteriophage coat protein and displayed on the surface of phages. Followed by repeated antigen or interacting protein selection and propagation, specific protein/antibody can be isolated and validated. This approach allows a rapid selection and large-scale production of specific proteins against potential binding partners. Compared to hybridoma technique limited to the fusion partner, phage display can apply to a broad range of species and discover the specific immunoglobulins. Available animals in Creative Biolabs including mouse, rat, rabbit, camel, llama, cattle, sheep, chicken, and others.
DNA Immunization and Immune Antibody Library Construction
As an alternative to conventional immunization method, scientists from Creative Biolabs are able to induce antigen-specific immune response through properly DNA immunization strategies. The immune antibody library can then be generated by our advance phage display platform to isolate novel antibodies against the naïve target.
Compared to the conventional method, DNA immunization takes advantages of expressing the interest antigen through the in vivo host cells. This approach overcomes the challenge of immunizing problematic proteins (e.g. membrane proteins, unstable proteins) and avoids the potential conformational variation caused by the recombinant proteins. Currently, Creative Biolabs offers a series of DNA immunization strategies to fit our clients’ diverse requirements.
On the basis of DNA immunized hosts, our scientists are pleased to utilize our phage display technology to transfer the valuable antibody repertoires to the immune library for further screening. To initiate library construction, lymphocytes are isolated from selected immunized animals, and followed by RNA isolation, cDNA generation, and PCR amplification. Encoding gene is packed with vector and introduced into host strains, such as E. coli, and undergo superinfection in phage display system (helper phages). Encode gene fuses with phage coat protein gene like pIII or pVIII and results in antibody assembling on the surface. These proteins/antibodies still retain their structure and attracted by ligand independently. Creative Biolabs provides multiple phage display systems including M13 phage system, T4 phage system, T7 phage system, and lambda phage system.
Biopanning and Screening
The diversity of generated phage libraries in Creative Biolabs ranged from 108 to 1012. Since phage display libraries contain a large number of phage particles and each individual has potential to encode protein ligands. It is necessary to conduct screening process that selects specific binders. Creative Biolabs offers biopanning service for the isolation of specific binders (antibodies). In biopanning, antigen or ligand binder are fixed on microtiter plate well, column matrix or magnetic beads as solid phase and phage particles are flowed as the mobile phase. Combined this system with repeated phage incubation, washing, and expansion, purified target binders can be eluted. Selected antibody display phage can be further evaluated in tissue section, ELISA, Western blotting, etc.
Through the integration of DNA immunization and phage display technique, Creative Biolabs is able to discover and develop naïve antigen specific antibodies from almost all species of experimental animals. Our seasoned scientists will provide the custom service to tailor the best fit immunization strategy and also the library construction and screening method to achieve our clients’ research objectives. To learn more details, please feel free to inquiry us.