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11.12 Tech Feat Antibods MH antibodies technology feature To affinity and beyond Antibodies are the researcher’s Swiss army knife: there’s a tool for every purpose, and they’re made in more shapes and sizes than ever. Pete Moore and Julie Clayton report. ntibodies not only protect us from from investors requires us to have a pretty For therapeutic use in humans, the gold infection, they have been exploited clear clinical focus.” standard is a fully human monoclonal anti- Afor years in the laboratory — in diag- Simple financial incentives drive much of body.Antibodies in which the antigen-bind- nostic tests, to purify proteins and as the the work in therapeutic antibodies. Donald ing site is from a mouse, even when most workhorses of cell biology to detect, locate Drakeman, president and chief executive of of the rest of the antibody is human, as in and identify cellular proteins. Designed by Medarex in Princeton, New Jersey, estimates chimaeric and humanized antibodies, will evolution to recognize and bind virtually that it takes some US$20 million to get a always risk triggering an unwanted immune any molecule that could exist, what more traditional small-molecule drug ready for response that, at best, neutralizes the thera- could we ask of them? Quite a lot, it seems. clinical testing, whereas this could cost only peutic antibody and renders it ineffective. To meet the demands of today’s researchers $2 million for an antibody. Finding the right human antibody has been and drug developers, antibodies are being made much simpler by the advent of genetic cut down to size, tweaked into new shapes, Consulting the library engineering and the development of vast and manufactured in entirely new ways. The business end of an antibody molecule human antibody libraries packed with Antibodies as research reagents are is the antigen-binding site. This is the part potential candidates. These libraries are ubiquitous. Therapeutic antibodies have had of the antibody that latches onto a specific based on phage-display technology and store a bumpier ride, but now they’re back — and antigenic determinant or epitope — a com- and express billions of human variable here to stay.Crucially,the large pharmaceuti- plementary structure on the surface of regions. Each phage carries a DNA sequence cal companies have regained interest. “Anti- another molecule. The antigen-binding site encoding a different variable region,and dis- bodies are causing a lot of excitement because is also known as the variable region, or vari- plays the corresponding protein on its sur- there are not many new drugs out there and able domain, as it is the part of the antibody face, usually as part of a phage coat protein. 30% of the biotech industry is driven by anti- molecule that varies most among the bil- When a potential drug target is screened body technology,” says Peter Hudson of lions of different antibodies that can be against the library, only phage carrying anti- CSIRO Health Sciences and Nutrition in made, either naturally in the human body bodies with an affinity for the target lock on Melbourne, Australia. Greg Winter of the or by protein engineering. As a large biolog- to it. The captured phage are multiplied in MRC Laboratory of Molecular Biology in ical molecule such as a protein can contain bacteria, and the antibody genes re-isolated, Cambridge, UK, and co-founder of antibody many epitopes, you can either produce to be fashioned into genes that can be used to drug-discovery company Domantis, agrees. monoclonal antibodies that target just one produce large amounts of fully functional “A year or two back we were interested in both epitope on a molecule, or raise a collection monoclonal antibodies in mammalian cell clinical and research applications, but it is of antibodies — polyclonal antibodies — cultures, or into smaller antibody fragments very difficult to make money from research that bind to a number of different epitopes that can be produced more cheaply in yeast applications and it is clear that the funding on the molecule. or bacterial culture. PLAYING WITH THE PIECES Regeneron of Tarryton, New York, has turned the usual way of using vascular endothelial growth factor (VEGF) is in clinical trials for the treatment antibodies as therapeutics on its head. To produce therapeutic agents that of cancer by preventing the growth of blood vessels into solid tumours and will mop up unwanted cytokines or growth factors, the company has non-Hodgkin’s lymphoma. “Every time we compare them with a developed a hybrid molecule called a ‘Trap’. This is made from the stem conventional antibody they have affinities two or three orders of magnitude of an antibody fused to two receptors for the target molecule. In this higher,” says George Yancopoulos, Regeneron’s chief scientific officer. application, the antibody stem Unlike antibodies, Traps will pick up every molecule that normally binds to gives the molecule a long half-life the receptor, so the VEGF Trap will also bind the related growth factor PLGF. in the circulation, while the Immunotoxins — antibodies linked to cytotoxic drugs — such as receptors replace the variable Wyeth’s Mylotarg, have the potential to avoid some of the toxic side effects regions. Traps therefore bind of traditional cancer chemotherapy by delivering the drug directly to the tightly to their target and can be tumour cell. Seattle Genetics of Washington state is developing SGN-15, given at infrequent intervals. an antibody–drug conjugate composed of a mouse–human chimaeric Regeneron has created an monoclonal antibody chemically linked to the cytotoxic drug doxorubicin. interleukin-1 (IL-1) Trap that it This is currently in a phase II clinical trial for the treatment of lung cancer. hopes may be effective against The antibody binds to a Lewisy-related antigen expressed on many types rheumatoid arthritis, and is of solid tumour. SGN-35, one of the company’s second-generation developing a dual IL-4/IL-13 Trap immunotoxins now in preclinical trials, incorporates a potent synthetic Regeneron’s VEGF Trap. as a potential treatment for asthma drug and a novel chemical linkage that makes the molecule more stable and other allergies. A Trap for in the blood. P.M. NATURE | VOL 426 | 11 DECEMBER 2003 | www.nature.com/nature © 2003 Nature Publishing Group 725 antibodies technology feature A recent success of the phage-display found in the blood of healthy peo- technology is HUMIRA, the first fully ple,but CAT has shuffled the DNA human monoclonal antibody to be approved sequences that code for the vari- for therapy, developed by Cambridge Anti- able region to give even more dif- body Technology (CAT) in Cambridge, UK, ferent types of antigen-binding and Abbott Laboratories in Abbott Park,Illi- site. Having started with human nois.This antibody,which blocks the inflam- genes, CAT believes its antibodies matory cytokine TNF-a,was approved in the are less likely to create immuno- United States in 2002 and in Europe in Sep- genic responses than libraries tember this year as a treatment for the symp- built with synthetic sequences. toms of rheumatoid arthritis. Antibody discovery and pro- CAT’s phage-display library holds single- duction company BioInvent in chain variable fragments (scFv) which con- Lund, Sweden, has a similar library sist of the variable ends of a light and a heavy called n-CoDeR, which holds chain linked with a short peptide.The library human scFv. Like CAT, BioInvent Going for GOLD with MorphoSys. now boasts more than 100 billion distinct starts with natural human antibody antigen-binding specificities. And in the genes as the source of the DNA sequences, cross-licensing its intellectual property with world of antibody discovery, most people which are then further diversified by making other major players, such as Genentech, think that size matters.The larger the library, variants of the antigen-binding regions. “We Xoma,Affimed,Biosite and CAT.Chief scien- the greater the chance that it contains an have taken evolution beyond nature and the tific officer Clive Wood believes that one of antibody for your given target.“It means you library now has more than 1010 members,”says the company’s strengths is the newness of its can get reasonably high-affinity antibodies Eskil Söderlind, who invented the n-CoDeR antibody libraries.“We are the new kid on the out of your first collection,”says Winter. technology with Carl Borrebaeck. BioInvent block, but because of that we have important The library started as a collection of anti- can screen up to 20,000 clones per day against a advantages.We have been able to incorporate body genes corresponding to antibodies customer’s target antigen on its automated new features into our Fab library,such as hav- system.The scFv fragment genes are then con- ing a mixture of natural and synthetic diversi- verted into complete antibody genes, and the ty that allows us to pull out antibodies with antibody itself produced in larger quantities picomolar to nanomolar affinity.”The largest BIOINVENT from genetically engineered mammalian cells. Dyax library contains about 37 billion dis- The biopharmaceutical company Dyax in tinct human antibodies, and the company Cambridge, Massachusetts, has phage-dis- has two therapeutic proteins derived from play libraries holding Fab fragments — an phage display in three phase II trials. entire single ‘arm’ of an antibody. Although The Human Combinatorial Antibody the company holds many of the foundation Library (HuCAL) of MorphoSys in Martin- patents for phage-display technology,Dyax is sried, Germany, offers 10 billion totally syn- a relative newcomer in applying phage dis- thetic variable regions modelled on partial play to antibodies,having initially focused on human variable regions and displayed in peptide and protein display.
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