Through a lot of hard work, a license from the Wisconsin Alumni Research Foundation (WARF) and a desire to have an impact on human health, NeoClone was founded in 1999 by three University of Wisconsin scientists who recognized the need for better ways to discover and develop antibodies. As they refined the process to make better antibodies (what is now the NeoAb process), they established a reagent catalog as well as a service business to make high-quality antibodies for others.
By 2002 the founders realized a need to bring in business expertise to help grow the company by leveraging its ability to make excellent antibodies. This change in direction is the foundation of what NeoClone is today - a company focused on developing high quality antibodies for sale as reagents and establishing partnerships and collaborations to develop diagnostics. As a result of these efforts, we now have over 1000 customers in 30+ countries.
NeoClone has always been a research-focused company with an emphasis on emerging antibody technologies. Reactivation is our proprietary technology for selecting, enriching, and amplifying murine or human antigen-specific antibody-producing cells. We stimulate only the class-switched (IgG), affinity matured Ig-secretors to proliferate, which increases the number of antigen -specific Ig-secreting cells relative to non-specific Ig-secreting cells.
To poise ourselves for future success, we've continued to refine our antibody development abilities. In 2014, we added antibody phage display technology to help us develop the diverse and specific antibody repertoire to certain difficult targets that might be difficult to achieve with hybridoma technology alone. We are now developing camelid (VHH) antibody capabilities so we can service the rapidly growing therapeutics market. We have had a research team focused on this effort and we will be offering these services in 2015.
NeoClone is a world-class custom antibody development company, having discovered hundreds of antibodies for our clients. But there are very difficult targets out there that hybridoma technology alone can’t solve. So we added antibody phage display capabilities to help us develop the diverse and specific antibody repertoire to difficult targets that you need. Our team now has over 15 years of scFv phage antibody and recombinant antibody experience.
Greater Diversity, Faster Answers
One of the advantages of phage display is the enormous diversity of variant antibody genes that can be represented in a phage library, even to relatively non-immunogenic targets. NeoClone’s antibody phage display service provides a means of rapidly screening large numbers of scFv in our antibody gene libraries against potential binding partners in only 2 weeks. Once promising clones are identified, we can clone, express and sequence the scFv antibodies for your further evaluation. Based on your needs, we can engineer the antibody further for expression in mammalian cells, affinity maturation or other optimization steps. NeoClone can also construct new scFv phage libraries from immunized or naïve donors from several species.
Antibody Phage Display Screening
NeoClone's proprietary antibody phage display screening method allows for efficient, high throughput identification of rare binders with distinctive characteristics required for a specific application. NeoClone is able to generate scFv antibodies that target antigens that have traditionally been difficult to attain, including Post-Translationally Modified peptides (PTMs) such as acetylated or phosphorylated antigens. In addition, NeoClone can express the specific phage as scFv-Fc in mammalian cell culture or by periplasmic expression in bacteria.
Most antibody developers only test if an antibody will recognize an antigen in an ELISA or a Western Blot assay. More often than not, however, these baseline assays may not provide you with the full range of information you need for a successful project. We believe that antibody developers should supply more, not less, information to clients.
That's why nearly all NeoClone projects involve multiple screenings, most of which are done in the early, discovery stage. Our extensive experience tells us that robust screening, at this point in the NeoAb process, will result in better focus on which clones to move into the development phase. Proper antibody characterization also helps determine the chances of overall project success.
What assays should be done in an antibody development project? Like much of the rest of the antibody development process, it depends on the target and desired end-use of the antibody. Of course you screen against the antigen, but you may also need to know:
These are but a few examples. Once we understand your specific needs, we can create a targeted approach to find the appropriate clones for development. At this point, we can work with you in one of two ways; we can either go into development with our selection of the best supernates, or we can send you the developed supernates so that you may complete your own screening while we complete ours. Many of our customers find the latter, collaborative approach to be the most effective.
In short, we believe that best way to develop great antibodies on a repeatable basis is by ensuring each antibody is fully characterized with robust screening.
Camelid VHH Antibody Services
NeoClone is fast becoming a leader in the single domain antibody library field. We offer a wide range of unique services, including construction of immunized single domain antibody libraries using our llamas, bio-panning of naïve or immunized single domain antibody libraries and large scale production of single domain antibodies (SdAb).
NeoClone's services include the generation and production of target-specific SdAb from either a naïve llama (camelid) VHH library or a custom immunized llama antibody phage display library. Following serial rounds of bio-panning against your target of interest, we use a proprietary selection procedure to identify the phage containing the target-specific VHH binding domain. The individual VHH antibody phage display genes that recognize your target are subcloned to provide multiple target-specific VHH antibodies in order to choose those with the highest affinity. From these isolated genes, the recombinant SdAb proteins are expressed and purified in vitro. Each recombinant VHH SdAb can be characterized and qualified for functionality in assays of your choice, including western blot, ELISA, immunoprecipitation, flow cytometry or immunohistochemistry.
Llama VHH (SdAb) background
The llama is a member of the camelidae family, which also includes alpacas and camels. In additional to conventional IgG, these animals produce subclasses IgG2 and IgG3, functional single chained antibodies that are capable of antigen binding. Referred to as VHH antibodies, these antibodies consist of only two heavy chains connected by several disulphide bonds in the hinge region. They lack the light chains and functional CH1 and CH4 domains. These heavy chain antibodies are being used to develop single domain antibodies. The single domain nature of these antibodies provide unique advantages which set them apart for research, diagnostic and therapeutic applications. Some of these advantages include:
A major roadblock to the development of immunotherapy has been the size of conventional antibodies; past a critical point the antibody is often unable to reach its target. The camelid heavy chain antibodies present advantages over common antibodies in the design, production, and application of clinically valuable reagents.
Screening our Naïve Llama VHH Library
NeoClone’s naïve camelid phage display single domain antibody library was developed from the PBMC of numerous llamas in order to broaden the diversity of the VHH genes generated in their combined immune responses. We can screen this library for your target quickly and move forward into developing a robust VHH antibody for you.
Custom Llama VHH Library Creation
We have upgraded our custom antibody capabilities to include the construction of immunized llama single domain antibody libraries that are based on phage display technology. In comparison with naïve libraries, immunized libraries usually produce a much more diverse panel of antibodies, and of even greater affinity, to your specific target of interest.
We are adept at cloning the single domain antibody repertoire from PBMC of immunized llamas into our phage display vectors. By reverse transcription and polymerase chain reaction, a library of single domain antibodies containing 10-100 million clones can be produced. This library will be enriched for your target, increasing the chances of getting just the right clone for your research, diagnostic or therapeutic need.
NeoClone has been developing high quality custom monoclonal antibodies for pharmaceutical, biotech, academic and federal laboratory customers for over 12 years. Using murine hybridoma technology, we have been quite successful at generating diverse panels of very high affinity monoclonal antibodies against difficult targets, such as:
Discovering great antibodies starts with generating a robust immune response against the antigen of interest. We understand and appreciate that every antigen is unique. Each project is discussed thoroughly before starting to be sure we’ll meet your needs. We also provide advice and technical expertise throughout the project to help you make the right decision at each step.
We believe that all successful projects start with great project design. This is where we excel. Using our NeoAb® bioinformatics approach, we help clients define a project that has the best chance of success in making antibodies that will meet their needs - in terms of both specificity and sensitivity.
In addition to some basic information needed from our clients (protein sequence, desired end use of the antibody), we help determine:
Based on the answers to these questions as well as others, we can design a project approach to minimize the inherent risk in antibody development, while at the same time increasing the chances of developing highly specific & sensitive antibodies of interest.
Because NeoClone is adept at both antibody phage display technology AND traditional hybridoma technology, we can help you decide which approach will work best for your needs.
Do you have a target of interest to which you would like NeoClone to apply its bioinformatic approach? Please contact us to begin the process.
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