Llama monoclonals production
Our optimized library construction and screening processes with phage display technology guarantee successful isolation of high affinity VHH clones in a short time.
Advantages of llama VHH single domain antibodies:
- VHH single domain antibodies are only 15kD in size, compared to about 150kD for typical immunoglobulins
- The smaller size of single domain antibodies allows them to physically fit into epitopes that are normally not accessible to traditional antibodies, including the active site of many enzymes
- The smaller size also enables them to penetrate tissues faster, even readily crossing the blood-brain barrier
- VHH antibodies are extremely stable and are resistant to both high acidity and temperature, even being able to fold back into a functional protein after denaturation
- Single domain antibodies have a very high solubility due to increased hydrophobicity
- The small size and stability makes the VHH antibodies ideal candidates for therapeutic and diagnostic applications.
Details of VHH single domain antibody production:
Llama immunization
- Phage display library construction: We monitor the anti-serum titer by ELISA and collect multiple production bleeds and PBMC cells throughout the immunization. The PBMC cells are preserved in RNA lysis buffer at different time point, so that the antibody repertoires are preserved through the immunization process. At the end of the immunization protocol, the client could choose the PBMC at any collection time point to construct the library.
- VHH library screening and affinity determination: The constructed library will be screened through multiple rounds of affinity binding/selection with antigen. Positive clones will be identified through ELISA. Genes of these positive clones will be sequence analyzed. Affinity of each binder will be determined through ELISA. Depending on the complexity of the antigen, one to three clones with the best affinity will be delivered.
- Large scale VHH antibody purification: For each positive clone, we will provide ~100ug of purified protein with no extra charge. Additional antibody protein are available at extra cost.
- Naive VHH phage display library: Our newest VHH library from non-immunized llamas covers large and diverse VHH repertoires representing over 2.0x109 peripheral mononuclear cells from 20 non-immunized llamas. The library has over 2x109 independent clones.