Abcore: A Renowned Name in Nanobody Discovery & Custom Antibody Production

Abcore is an industry leader in nanobody discovery and custom antibody production, located in sunny Southern California, USA. The local climate is mild year-round, which allows the animals to be housed in "open-air" facilities. This natural environment, along with constant attention to our animals' health and comfort, leads to a better immune response and higher-quality antibody production. Abcore has developed a complete nanobody platform based on antibody discovery via phage display and has made a variety of native phage display libraries that we can use for animal-free antibody discovery. Abcore offers a wide variety of polyclonal, monoclonal, and recombinant antibodies and is the largest camelid facility in the United States. Abcore is an established CRO and has worked with some of the world's largest biotechnology and pharmaceutical companies.

BTL Biotechno Labs Pvt. Ltd. is an honoured channel partner of Abcore and offers their complete range of services, including Nanobody Discovery, Custom Monoclonal & Polyclonal Antibody Production and CRO services such as Nanobody Phage Display, Phage Library Panning & Screening, Nanobody Expression, Purification & Humanization for extensive research at the proteomic, genomic, and molecular levels in India. By combining our expertise and resources, we are bringing cutting-edge nanotechnology-enabled products to the Indian market.

Abcore is an established CRO with a platform for antibody discovery and nanobody discovery. They work with some of the largest biotechnology and pharmaceutical companies in the world. They have a full platform of services based around single-domain antibody discovery and nanobody characterization.

CRO Services:

• Nanobody Discovery
• Nanobody Phage Display
• Phage Library Panning
• Phage Library Screening
• Nanobody Expression and Purification
• Nanobody Humanization
• Nanobody Cell Binding
• Generation of Naïve Phage Libraries
• Generation of Immunized Phage Libraries

Abcore has an entire platform built around discovery and characterization of VHH nanobodies. They have a farm with over 300 llamas and 150 alpacas and have highly characterized naïve phage libraries. They have optimized processes for generation of these libraries using proprietary primers that have higher coverage than the traditional published primers. Once positive VHH sequences are discovered, Abcore can purify the nanobodies, characterize the nanobodies, and optimize the nanobodies for various experimental and clinical applications. Abcore has built an entire platform for nanobody and single domain antibody discovery. Their process is based on phage display and using either Naïve phage libraries or immunized phage libraries for nanobody discovery.

Immunized Antibody Phage Libraries:

• Custom Llama Nanobody Discovery
• Custom Alpaca Nanobody Discovery

Naïve Antibody Phage Libraries:

• Naïve Llama Nanobody Phage Library
• Naïve Alpaca Nanobody Phage Library
• Naïve Camel Nanobody Phage Library
• Naïve Shark IgNAR Phage Library
• Naïve Rabbit ScFv Phage Library

The nanobody discovery process can be broken into phases, based on the goals of the single domain antibody project:

1. Immunization
2. BMC Isolation
3. cDNA generation
4. Phage Library Construction
5. Phage Library Panning
6. Screening for Positive Clones
7. Production of Purified Nanobodies

Abcore has developed proprietary procedures for nanobody discovery. Their optimized library construction and screening processes with phage display technology guarantee successful isolation of high affinity llama nanobodies in a short time. They have successfully produced multiple positive clones for a variety of antigens, ranging from large protein molecules (>400kD) to small haptens (small chemical molecules of ~200 dalton), many of the binders have sub-nanomolar affinities measured by ELISA.

Details of VHH single domain antibody production

• Llama immunization: Animal immunization is handled by Abcore exclusively. Abcore is well established for llama immunization.
• Phage display library construction: They 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 panning and screening: 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.

Nanobody Phage Display

One of the most important parts of Abcore’s nanobody discovery platform is their proprietary phage display technology. Their phage display starts with either an immunized or naive nanobody library. They first use panning to enrich the library for nanobodies that bind to the target. They often use pre-panning steps to address the cross-reactivity goals of the project. They can pan to remove any binders to closely related isoforms or use a parental or knock-out cell line for cell-based projects. After the rounds of screening, and with good enrichment, they move on to selection. Abcore’s proprietary OPIS technology allows it to screen the enriched phage pools for positive binders in the absence of phage. This removes the possibility of non-specific phage binders (phage is known for being very sticky) and only detects nanobodies that bind to the target of interest. Abcore actually confirms binding to the target twice before even sequencing the clones.

Nanobody Expression and Purification

Abcore’s antibody discovery platform ends with the delivery of positive, unique nanobody sequences. Often, the next step in the process is to produce the nanobody for further testing. Depending on what further characterizations of the nanobody protein will be done, proteins can be produced in a few different purities. Currently, all the protein expression is done in E. coli. The first step is to grow a culture in volumes ranging from 5 ml up to multiple litres. Then E. coli undergoes osmotic shock, where the nanobodies can be isolated from the periplasmic space. This "shockate" is about 80–90% pure and is pure enough for many further experiments. If the nanobody needs to be more pure or the concentrations need to be normalised for further experimentation, it can be further purified. In this case, Abcore purifies the nanobodies that have a His tag, and we can purify them using Ni-NTA (Nickel-NTA) chromatography. This results in a highly pure, known concentration of purified nanobody protein.

Nanobody Humanization

Abcore can humanize nanobodies for clinical applications. Humanization of the VHH region of nanobodies is fairly straightforward and something we do often for our pharma companies. If a nanobody is headed for the clinic, this is an integral step. Abcore will always strive to achieve full humanization of a VHH nanobody. Sometimes full humanization is possible, but other times modification of too many of the amino acids will result in instability of the nanobody, resulting in stability or solubility problems. Abcore will generate multiple sequences, from the native VHH sequence to fully humanized, and then test the proteins for stability and also make sure that they maintain binding to the target of interest. Once the nanobody is humanized, it can undergo further modifications for clinical applications.

Nanobodies have numerous beneficial properties, such as their small size, which allows them easier access to tissues; high stability; and a strong affinity for their cognate antigen, all of which make them very good candidates for development into next-generation biopharmaceutical agents. BTL Biotechno Labs Pvt. Ltd., being a partner of Abcore, provides their Nanobody custom services along with technical support in India at an affordable price.

Other CRO Services (Humanization, Functional Assays, Expression Vector changes, etc.) Abcore is an industry leader in custom antibody production, and their service offerings continue to grow. They have completed tens of thousands of traditional polyclonal antibody projects, and offer polyclonal antibody production in rabbits, goats, and sheep. They also perform ascites production in mice. Rabbits are the normal go to animal for production of antibodies because of their size, ease of handling, and wide array of secondary antibodies that exist.

Polyclonal Antibody Services:

• Rabbit Polyclonal Antibody Production
• Goat Polyclonal Antibody Production
• Sheep Polyclonal Antibody Production

Monoclonal Antibody Services:

• Ascites Production

We, BTL Biotechno Labs Pvt. Ltd., a distributor of Abcore in India, provide their complete range of quality custom services such as Nanobody Discovery, Custom Monoclonal & Polyclonal Antibody Production and CRO services such as Nanobody Phage Display, Phage Library Panning & Screening, Nanobody Expression, Purification & Humanization to promote immunology research at the best price with proper technical support.

For more details, please follow the link below: https://abcore.com