Select Suitable Immunization Strategies for VHH Discovery
Multiple Sites Immunization DNA Immunization Whole Cell Immunization Case Study Published Data FAQ Resources
As a pioneer and undisputed global leader in the field of VHH development, Creative Biolabs is proficient in applying advanced immunization strategies to satisfy various project demands. Based on our extensive experience, we have earned a great reputation among our worldwide clients for successfully developing a series of high-quality VHHs against various challenging targets, such as small molecules, membrane proteins, and PTM-modified targets.
Depending on the target of interest and the specific project objective, our scientists can tailor the best-fit strategies to establish an appropriate immunization program. This program will involve one of the following typical strategies or combine some of them as a heterologous strategy, which can contribute to triggering the best immune response.
Multiple Sites Immunization
Multiple sites immunization is the most standard strategy for large animal immunization. This strategy can elicit responses from several easily accessible sites, thereby contributing to the development of antibodies with high specificity and affinity. It is also the best option if the injection of more than one antigen is required.
Typical Advantages of Multiple Sites Immunization
- Triggers the desired immune response with a smaller amount of antigen
- Allows the involvement of different antigens in a single immunization round
- Compatible with various antigen formats
DNA Immunization
DNA immunization is an efficient approach to eliciting an immune response against challenging antigens. Compared with traditional immunization methods, DNA immunization allows for antigen production in vivo, thus avoiding the time-consuming processes of antigen preparation. What's more, the naïve structure of the expressed antigens permits the generation of antibodies with high specificity and affinity.
Typical Advantages of DNA Immunization
- Available for challenging antigens, such as insoluble proteins, multi-pass membrane proteins, secreted proteins, intracellular proteins, unstable proteins, etc.
- Time- and labor-saving
- Avoids the risk of antigen contamination
- Generates antibodies with high affinity and specificity
Whole Cell Immunization
Whole cell immunization is a novel strategy to elicit an immune response against an antigen over-expressing cell line. Unlike conventional immunization strategies, whole cell immunization benefits from higher immunogenicity, naïve antigen structure, and a simpler purification process. It is one of the best methods to discover novel VHHs against the extracellular domain of desired membrane protein targets.
Typical Advantages of Whole Cell Immunization
- Allows the discovery of antibodies against the extracellular domain of membrane proteins
- A powerful tool for identifying new biomarkers from surface proteins of tumor cell lines
- Enables the generation of novel antibodies that recognize the naïve conformation of a target
With over a decade of extensive experience in providing immune-based VHH discovery projects, Creative Biolabs has contributed to discovering thousands of novel single domain antibodies for customers worldwide. If you are interested in our services, please do not hesitate to contact us for more details.
Published Data
- An Efficient Genetic Immunization Strategy for Generating VHHs
Fig. 1 Proposed mechanism of heavy-chain antibody induction by genetic immunization.1,2
Fig. 2 CHO cells were transiently co-transfected with expression vectors to identify
specific binders.1,2
This article introduces an efficient genetic immunization strategy for generating VHHs against membrane proteins in their native conformation. As illustrated in Fig. 1, the study outlines a genetic immunization strategy that includes delivering cDNA plasmids using gene gun technology, in vivo transcription of cDNA into mRNA, and subsequent translation into target membrane proteins, which induces B cells to produce specific VHHs. It has been demonstrated that this genetic immunization strategy can effectively produce the desired membrane proteins in host cells and activate specific immune responses to produce high-affinity antibodies through a process of somatic hypermutation. Fig. 2 shows an effective high-throughput screening method that uses VHH-based heavy chain antibodies for cell surface binding screening and detection with fluorescein-labeled secondary antibodies. This method can be utilized to identify VHHs targeting specific membrane proteins, thereby accelerating the discovery and optimization of VHHs, which is crucial for developing novel diagnostic and therapeutic tools.
References
- Eden, Thomas, et al. "A cDNA immunization strategy to generate nanobodies against membrane proteins in native conformation." Frontiers in immunology 8 (2018): 1989.
- Under Open Access license CC BY 4.0, without modification.
FAQ
1. Q: What is the general immunization timeline?
2. Q: How many animals will be used for each project, any recommendation?
3. Q: What will happen if the animal died due to the toxic antigen?
4. Q: Is that available to keep the host animal after immunization?
5. Q: What are the acceptable antigen types for immunization?
6. Q: What is the least antigen quantity for an immunization procedure?
7. Q: What's the difference between camel, llama, and alpaca?
8. Q: What is required for DNA immunization and whole cell immunization?
9. Q: Deliverables of Immunization Stage
Resources
Case Study
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