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U-READ - Unit for Rapid Engineered Antibody Development

The continuous demand from basic and applied research is increasing the need for isolating antibodies with desired properties in more efficient manners. Recombinant antibody technologies combined with in vitro display methods, such as phage display, offers a great alternative to traditional immunization techniques. Not only does it provide a renewable monoclonal source of antibodies, the process generates characterized antibody fragments in approximately three to four weeks compared to months with immunizations. Phage display technology allows control over the selection processes to a higher degree than what is possible with immunizations. Also, since the system allows the DNA encoding the antibody fragments to be easily manipulated, fine-tuning of affinity and specificity can easily be done. In addition, since all the steps of the selection and screening process are liquid based they can be automated. 

At the department we have set-up a high-throughput phage selection and screening pipeline that allows us to generate binders to a large number of antigens in a short time frame. This pipeline was originally scaled-up as part of AFFINOMICS, an EC FP7 Collaborative Project.

The Unit is able to select specific antibodies from large in-house developed antibody libraries using phage display technology. If you need more information on such efforts or are interested in using U-READ for your research efforts, please contact Mats Ohlin. The unit is also part of SciLifeLab's Human Antibody Therapeutics National Facility within its Drug Discovery and Development platform. This National Facility supports the development of therapeutic antibodies to targets as suggested by the partner primary investigators.

As proof of principle, generated antibodies are used in-house in a variety of research applications, for instance in our various affinity proteomics platforms.
  
  

Technical platforms

The unit uses its in-house developed antibody fragment (scFv) library for selection of specific antibodies using phage display technology. We host a variety of technology platforms such as

  • Hamilton robot for efficient liquid handling
  • King Fisher robot for efficient, parallell phage display selection procedures
  • a variety of immunoassay platforms including ELISA, Luminex etc.
  • MASS-1 surface plasmon resonance-based real time high through-put interaction assessment platform
  • iQue high content, high throughput flow cytometry assay platform for analysis of protein binding to cells and particles.

The Department also host a variety of other technology platforms including flow cytometry (analyzer as well as cell sorter), qPCT, nanostring nCounter analysis system, Affimetrix microarrays, etc.
   

References

  1. Ljungars A, Svensson C, Carlsson A, Birgersson E, Tornberg U-C, Frendéus B, Ohlin M, Mattsson M (2019) Deep mining of complex antibody phage pools generated by cell panning enables discovery of rare antibodies binding new targets and epitopes. Front Pharmacol (in press) (Abstract)
  2. Ljungars A, Mårtensson L, Mattsson J, Kovacek M, Sundberg A, Tornberg UC, Jansson B, Persson N, Kuci Emruli V, Ek S, Jerkeman M, Hansson M, Juliusson G, Ohlin M, Frendéus B, Teige I, Mattsson M (2018) Phenotypic discovery of therapeutic antibodies and targets for cancer treatment. NPJ Precis Oncol 2, 18. (Abstract)
  3. Persson H, Kirik U, Thörnqvist L, Greiff L, Levander F, Ohlin M (2018) In vitro evolution of antibodies inspired by in vivo evolution. Front Immunol 9, 1391. (Abstract)
  4. Säll A, Walle M, Wingren C, Müller S, Nyman T, Vala A, Ohlin M, Borrebaeck CAK, Persson H (2016) Generation and analyses of human synthetic antibody libraries and their application for protein microarrays. Prot Eng Des Select 29, 427-437. (Abstract)
  5. Säll A, Persson H, Ohlin M, Borrebaeck CAK, Wingren C (2015) Advancing the Global Proteome Survey platform by using an oriented single chain antibody fragment immobilization approach. New Biotechnol 33, 503-513. (Abstract)
  6. Säll A, Sjöholm K, Waldemarson S, Happonen L, Karlsson C, Persson H, Malmström J (2015) Development of phage-based antibody fragment reagents for affinity enrichment of bacterial IgG binding proteins. J Proteome Res 14,4704-4713. (Abstract)
  7. Gerdtsson AS, Malats N, Säll A, Real FX, Porta M, Skoog P, Persson H, Wingren C, Borrebaeck CAK (2015) A multicenter trial defining a serum protein signature associated with pancreatic ductal adenocarcinoma. Int J Proteomics 2015, 587250. (Abstract)
  8. Säll A, Carlsson F, Olsson N, Wingren C, Ohlin M, Persson H, Waldemarson S (2014) AFFIRM – a multiplexed immunoaffinity platform that combines recombinant antibody fragments and LC-SRM analysis. J Proteome Res 13, 5837-5847. (Abstract)
  9. Taussig MJ, Stoevesandt O, Borrebaeck CAK, Bradbury AR, Cahill D, Cambillau C, de Daruvar A, Dübel S, Eichler J, Frank R, Gibson TJ, Gloriam D, Gold L, Herberg FW, Hermjakob H, Hoheisel JD, Joos TO, Kallioniemi O, Koegll M, Konthur Z, Korn B, Kremmer E, Krobitsch S, Landegren U, van der Maarel S, McCafferty J, Muyldermans S, Nygren PÅ, Palcy S, Plückthun A, Polic B, Przybylski M, Saviranta P, Sawyer A, Sherman DJ, Skerra A, Templin M, Ueffing M, Uhlén M (2007) ProteomeBinders: planning a European resource of affinity reagents for analysis of the human proteome. Nat Methods 4, 13-17. (Abstract)