Featured resource: Developmental Studies Hybridoma Bank
Posted by Douglas Houston, on 31 January 2022
Doing great science depends on teamwork, whether this is within the lab or in collaboration with other labs. However, sometimes the resources that support our work can be overlooked. In our new series, we aim to shine a light on these unsung heroes of the science world. The fourth article in the series is by Doug Houston (Interim Director, DSHB) who describes the work of the Developmental Studies Hybridoma Bank.
A short history of monoclonal antibodies and the DSHB
In 1975, Köhler and Milstein1 published a method for selecting and immortalizing cells that secreted a single, monospecific antibody, or monoclonal antibody. This advance ushered in the era of ‘biologics’, or biological molecules with exquisite specificity akin to chemical molecules. Monoclonal antibodies are made by fusing an antibody-producing mature B cell (usually obtained from the spleen of an immunized mouse) and an immortalized myeloma cell, producing a ‘hybridoma”. As the number of labs making and using these hybridomas grew, it became apparent that community repositories would be needed to effectively preserve and share these reagents.
The DSHB was one of these early repositories, and is one of the few, if not the only one, still in existence and operating independently. The DSHB was formed in 1986 through a contract with the National Institute of Child Health and Human Development (NICHD) involving Thomas August at Johns Hopkins University and Michael Solursh at the University of Iowa. Dr. August ran the Johns Hopkins portion, which generated new hybridomas, mainly focusing on immune cell development, while the University of Iowa group directed by Prof. Solursh in the Department of Biology stored and distributed hybridomas and antibodies.
Prof. Solursh’s lab was focused on limb development and early hybridoma deposits reflected these interests. Early hybridoma deposits expressed antibodies recognizing important molecules in limb development including muscle and matrix proteins. One of the first antibodies deposited was MF 20 (anti-sarcomeric myosin heavy chain), which was contributed by Donald Fischman in 1986. MF 20 remains one of the most requested and robust monoclonal antibodies in the DSHB collection. The skeletal muscle and extracellular matrix hybridoma collections grew substantially in the initial years. After Prof. Solursh tragically died at a young age in 1994, the Iowa division of the DSHB came under the interim directorship of David Soll, and daily operations were run by Karen Jensen, a long-time research associate in the Solursh Lab. As the collection of hybridomas in Iowa grew and the depth of curation and expertise surrounding these antibodies became established, the DSHB was consolidated at Iowa under Prof. Soll’s directorship.
In 1998, the NICHD’s contract with the DSHB was ended (three years ahead of schedule) and the Bank has been independent of NIH and self-supported ever since. Our operating expenses are covered by distribution fees, which are kept as low as possible (at cost) to facilitate wide use of the antibodies/hybridomas by the worldwide scientific research community. The DSHB still maintains close relationships with various NIH entities, including the National Cancer Institute (NCI) and the Common Fund. The DSHB is the preferred distributor for NIH, HHMI and MDA funded monoclonal antibody efforts (among others).
The people behind the DSHB
The DSHB is run by a relatively small but dedicated group of office staff and scientists. Our staff are experts at navigating the changing rules of university policies, shipping and customs rules, and addressing the concerns of researchers from around the globe. Many have been with the Bank for over a decade and a select few have worked with the DSHB since its inception. Notably, Dr. Karla Daniels (the voice on the phone when you call us!) worked with Profs. Solursh and Soll and has defined the role of our senior scientific curator of the DSHB collection and expert technical advisor, disseminating information on best use of our antibodies. Brian and Rebecca maintain the hybridomas cell lines and produce the antibodies we distribute. Working with hybridomas is an artisanal science, and Brian and Rebecca have mastered the art of coaxing these cells out of cryostorage and getting them to produce large quantities of immunoglobins. Nicki, Mitch and Brian process and manage the orders and track them until delivered to the destination labs. We’ve recently hired a dynamic new cohort of staff including Nick, Mejd and Nisha to continue the tradition of high-quality antibody production and service
What is available for researchers?
The DSHB provides two valuable services for researchers. First, scientists can ‘deposit’ their hybridomas (or antibodies) with the DSHB, granting the DSHB non-exclusive rights to distribute the antibody products as well the hybridoma cell lines (if desired) to the scientific research community. The depositor and the originating institution retain the intellectual property to the hybridoma and thus can benefit from commercialization while still making the reagent available to researchers. Second, researchers can purchase antibodies contributed by other scientists, with the stipulations that the reagents be used for non-commercial research or teaching and not be transferred or reverse engineered. The original mandate of the DSHB was to keep the costs to researchers as low as possible, and we continually strive to be the main provider of high-quality but affordable antibodies. Researchers typically purchase hybridoma-conditioned cell culture supernatant to use in immunostaining or immunoblotting, although other higher concentration options such as bioreactor supernatants or concentrated supernatants are available. Custom orders for large amounts of antibody or purified immunoglobin may also be requested.
We also accept deposits of polyclonal antisera, and although it has not been widely advertised (yet!), the DSHB is also accepting plasmids encoding recombinant antibodies and distributing the antibodies produced by transfected cells (DSHB will not distribute the plasmids themselves). These recombinant antibodies are an important recent advance in antibody technology and are the main way to produce monoclonal antibodies made in rabbits. Rabbit antibodies (traditionally used as polyclonal antisera) have different properties from those of mice, including better responses to shorter epitopes and a more diverse immune response2. Rabbits may thus be better for making antibodies against phospho- or other modified amino acid epitopes and represent an important complement to traditional mouse monoclonals.
How can the community contribute?
Because the DSHB considers itself a community resource, the research community provides critical feedback and support for the DSHB. Scientists can help support the DSHB by contributing hybridomas and/or antibodies when appropriate. It’s humbling to see the list of prominent scientists who have contributed reagents (see the ‘contributor spotlight’ on the DSHB website: https://dshb.biology.uiowa.edu). Also, the DSHB and other repositories can be supported by purchasing from them whenever possible. The DSHB and other repositories are critical for fostering rigor and reproducibility in research, a goal promoted by NIH and other funding agencies. Depositing with the DSHB is thus an easy way to implement ‘resource sharing plans’ in grant applications.
Because part of our mission is to keep costs low, the DSHB (unfortunately) does not actively characterize or validate each antibody, but depend on the depositor’s validation. Validation builds as each antibody accrues citations and confirming applications. Users can contribute by sending feedback about antibodies received from the DSHB. The low cost of DSHB is advantageous for experimenting with different conditions and antibodies, and we hope that labs tell us what works and what doesn’t! As community efforts directed at validating monoclonal antibodies improve (do they specifically recognize the target antigen, and only the target antigen?), we hope to improve access to this information for our collection. Users can always send us images of the antibodies at work, and if we can’t use them on the web site, we’ll disseminate on social media (Twitter: @_DSHB_; FB: https://www.facebook.com/DSHBUI/).
And last, as always, please cite the antibodies and contributors of useful antibodies. This is especially important for new or untried antibodies; many of these are potentially useful but may not be widely adopted unless cited in the literature.
User comments
Many antibodies at the DSHB are unique, absent from the catalogues of large antibody companies, and we often hear from customers how antibodies in the DSHB collection are essential to their research. This is especially true for those working in model organisms where commonly available antibodies made against human and mouse homologues may not work. Many commenters wish we had more antibodies in our collection. We also receive comments appreciative of our customer service. Of course, we receive the ‘rare’ complaint too. We actively troubleshoot use of the antibodies, drawing on the expertise of our scientists, who have not only deep general knowledge, but often long histories of working specifically with the antibodies in question over the years. Our favourite comment however is from someone who wished we were a hundred times bigger!
We hold this as an aspirational goal. If money were no object, we would exponentially grow the number of antibodies to human proteins and establish a robust validation pipeline. But regardless, the DSHB will continue to foster the development and sharing of antibodies for model (and non-model) organisms, which have contributed the most to the discovery of new biological insights.
Any hidden gems, features that are new, or that researchers might be less aware of?
The DSHB collection has continued to grow, from several hundred in the 1980s -1990s to over 5000 currently. Recent additions include a large number of recombinant antibodies from the Clinical Proteomic Technologies for Cancer program of the National Cancer Institute, which includes over 800 antibodies against cancer-related targets. These include many antibodies related to DNA damage repair, RAS/MAPK signalling, including regulatory phospho-epitopes, as well many transcription factors involved in epithelial-mesenchymal transition. Through the Protein Capture Reagent program of the NIH Director’s Common Fund, we took deposit of over 700 transcription factor antibodies, optimized for ChIP3. These were recently evaluated for ChIP-exo/seq by Frank Pugh’s group at Penn State University, in a recent publication, and many antibodies gave superior results using just ‘raw’ antibody-containing supernatant4. Also, last year, the NeuroMab collection of neuroscience and neurodevelopment antibodies (over 500 hybridomas), developed by James Trimmer at UC Davis5, was deposited with the DSHB. There are doubtless many hidden gems among the uncharacterized antibodies at the DSHB, although we think the DSHB itself is also a hidden gem, and we are grateful to the Node for featuring us and other science resources!
Douglas W. Houston
Interim Director, DSHB
Iowa City, IA. Jan. 2022
1Köhler, G. and Milstein, C. (1975). Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256, 495–497.
2Weber, J., Peng, H. and Rader, C. (2017). From rabbit antibody repertoires to rabbit monoclonal antibodies. Exp Mol Medicine 49, e305.
3Venkataraman, A., Yang, K., Irizarry, J., Mackiewicz, M., Mita, P., Kuang, Z., Xue, L., Ghosh, D., Liu, S., Ramos, P., et al. (2018). A toolbox of immunoprecipitation-grade monoclonal antibodies to human transcription factors. Nature methods 15, 330–338.
4Lai, W. K. M., Mariani, L., Rothschild, G., Smith, E. R., Venters, B. J., Blanda, T. R., Kuntala, P. K., Bocklund, K., Mairose, J., Dweikat, S. N., et al. (2021). A ChIP-exo screen of 887 PCRP transcription factor antibodies in human cells. Genome Research 31:1663-1679.
5Gong, B., Murray, K. D. and Trimmer, J. S. (2016). Developing high-quality mouse monoclonal antibodies for neuroscience research – approaches, perspectives and opportunities. New Biotechnol 33, 551–564
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