Nanobody immunized library
Our typical workflow for the discovery of nanobodies using an immunized library involves several key stages:
The first step involves the immunization of camelids or sharks with the antigen of interest. We work closely with our clients to design an optimized immunization protocol, tailored to their specific antigen of interest. We carefully select the most appropriate animal species and immunization schedule to ensure a robust and specific immune response.
Once the animal has been immunized, we collect peripheral blood mononuclear cells (PBMCs) from the animal. PBMCs are isolated from whole blood using standard laboratory techniques and contain the immune cells, including B cells, which produce the antibodies of interest.
Next, we use the PBMCs collected from the animal to generate a library of nanobodies. We use state-of-the-art technologies and techniques to construct a highly diverse and representative library of single-domain antibodies. This involves isolating the mRNA from the immune cells and converting it to cDNA, which is then amplified using PCR. The amplified DNA is then cloned into a vector to create a library of nanobodies.
Once the library has been constructed, we employ high-throughput screening techniques to identify nanobodies with high affinity and specificity toward the target antigen. We use advanced methods such as phage display to rapidly screen the library for binders and perform multiple rounds of screening and optimization to further enhance the binding properties of the selected nanobodies.
Nanobody production and purification
Once the desired nanobodies have been identified, we produce them in large quantities using recombinant protein expression systems. This involves cloning the selected antibodies into expression vectors and transfecting them into host cells. The antibodies are then purified to high levels of purity using standard laboratory techniques.
Finally, we validate the selected nanobodies for their binding specificity and functionality. We use a range of methods, including ELISA, Western blotting, and functional assays, to confirm the binding properties of the antibodies. We work closely with our clients to ensure that the selected antibodies meet their specific needs and requirements.
Throughout the workflow, our team of experts works closely with our clients to ensure their specific needs and requirements are met. We provide regular updates on the progress of the project and are committed to delivering high-quality and reliable results in a timely manner.
Turnaround time for discovery services
|I||Antigen preparation||TBD||QC reports|
~10 weeks (Camelids)
|Sera, PBMC, antibody titer|
|III||Immune phage display library construction||~3 weeks||Immune library construction, library quality evaluation|
|IV||Antigen-specific binder screening||~3 weeks||Enriched antigen-specific phage after 3~4 rounds of panning|
|V||Binder identification and validation||~3 weeks||Sequences of positive clones, purified binders, bioactivity validation|
Data analysis and report writing
|~1 week||Project final report|
5~6 months (Camelids)