Milstein and Kohler: Monoclonal Antibodies.

César Milstein (1927-2002) was born in Bahía Blanca, Argentina. He was a chemistry student educated at the University of Buenos Aires, obtaining his degree in biochemistry in 1952. He completed his PhD in 1957 at the same University and also received the British Council research scholarship to the biochemistry department at the University of Cambridge. Under the direction of the distinguished biochemist Frederick Sanger, Milstein completed a second dissertation, a study of the enzyme, earning his PhD in 1960 and joining the scientific staff of the Medical Research Council (MRC). He then worked at the National Institute of Microbiology in Buenos Aires from 1961 to 1963 until political turmoil led him to resign and return to Cambridge to work with Sanger at the newly formed MRC Laboratory of Molecular Biology. Now, he shifted his focus from enzymes to antibodies. By the early 1970s, he was an internationally recognized leader in the field and was able to attract talented, young researchers, including Georges Köhler, to the MRC. In 1983, Milstein was named head of the Protein and Nucleic Acid Chemistry Division of the MRC, a position he held until his retirement in 1995. He died in Cambridge, England, in 2002 at the age of 74.

[Humanization of Murine Monoclonal anti-hTNF Antibody: The F10 Story].

Tumor necrosis factor (TNF) is a proinflammatory cytokine implicated in pathogenesis of multiple autoimmune and inflammatory diseases. Anti-TNF therapy has revolutionized the therapeutic paradigms of autoimmune diseases and became one of the most successful examples of the clinical use of monoclonal antibodies. Currently, anti-TNF therapy is used by millions of patients worldwide. At the moment, fully human anti-TNF antibody Adalimumab is the best-selling anti-cytokine drug in the world. Here, we present a story about a highly potent anti-TNF monoclonal antibody initially characterized more than 20 years ago and further developed into chimeric and humanized versions. We present comparative analysis of this antibody with Infliximab and Adalimumab.

Is tumor necrosis factor-α friend or foe for chronic heart failure?

Although detrimental effects of tumor necrosis factor-α (TNF-α) have been reported in failing myocardium, clinical trials using TNF-α antagonists did not show the benefit of TNF-α inhibition in patients with chronic heart failure (CHF). The double-edged effects of TNF-α/Toll-like receptors (TLRs)-related proinflammatory cytokines and downstream signal transduction, nuclear factor (NF)-κB activation on failing myocardium are discussed.

Development of anti-TNF therapy for rheumatoid arthritis.

The aetiology of systemic, autoimmune, chronic inflammatory diseases--such as rheumatoid arthritis--is not known, and their pathogenesis is complex and multifactorial. However, progress in the characterization of intercellular mediators--proteins that are now known as cytokines--has led to the realization that one cytokine, tumour-necrosis factor (TNF; previously known as TNF-alpha), has an important role in the pathogenesis of rheumatoid arthritis. This discovery heralded a new era of targeted and highly effective therapeutics for rheumatoid arthritis and, subsequently, other chronic inflammatory diseases.

The first decade of biologic TNF antagonists in clinical practice: lessons learned, unresolved issues and future directions.

Results from clinical trials of biologic anti-TNF drugs performed in the late 1990s confirmed the biological relevance of TNF function in the pathogenesis of chronic noninfectious inflammation of joints, skin and gut, which collectively affects 2-3% of the population. Up to April 2009, more than two million patients worldwide have received the first marketed drugs, namely the monoclonal anti-TNF antibodies infliximab and adalimumab and the soluble TNF receptor etanercept. All three are equally effective in rheumatoid arthritis, ankylosing spondylitis, psoriasis and psoriatic arthritis, but, for not clearly defined reasons, only the monoclonal antibodies are effective in inflammatory bowel disease. About 60% of patients who do not benefit from standard nonbiologic treatments for these diseases respond to TNF antagonists. Less than half of responding patients achieve complete remission of disease. Importantly, some of those patients with rheumatoid arthritis in whom long-term anti-TNF therapy induced disease remission remain disease-free after discontinuation of any kind of treatment. There are not yet reliable predictors of which patients will or will not respond on anti-TNF therapy, whereas subsequent loss of an initial clinical response occurs frequently. The spectrum of efficacy anti-TNF therapies widens to include diseases such as systemic vasculitis and sight-threatening uveitis. While paradoxical new adverse effects are recognized, i.e. exacerbation or development of new onset psoriasis, reactivation of latent tuberculosis remains the most important safety issue of anti-TNF therapies. Clinical practice guidelines and consensus statements on the criteria of introduction, duration of treatment and cessation of TNF antagonists, including safety issues, are under constant revision as data from longer periods of patient exposure accumulate. It is hoped that more efficacious drugs that will ideally target the deleterious proinflammatory properties of TNF without compromising its protective role in host defense and (auto)immunity will be available in the near future.

[A short history of anti-rheumatic therapy--VII. Biological agents]. / Piccola storia della terapia antireumatica--VII. I farmaci "biologici".

The introduction of biological agents has been a major turning-point in the treatment of rheumatic diseases, particularly in rheumatoid arthritis. This review describes the principle milestones that have led, through the knowledge of the structure and functions of nucleic acids, to the development of production techniques of the three major families of biological agents: proteins, monoclonal antibodies and fusion proteins. A brief history has also been traced of the cytokines most involved in the pathogenesis of inflammatory rheumatic diseases (IL-1 and TNF) and the steps which have led to the use of the main biological drugs in rheumatology: anakinra, infliximab, adalimumab, etanercept and rituximab.

The history of the antibody as a tool.

Antibodies are essential tools in modern science and medicine, however the history leading to the use of antibodies as tools has not been well-described. The objective of this paper is to analyze the history of immunology from smallpox inoculation to the production of monoclonal antibodies, and to identify turning points in immunological theory leading to the emergence of antibody-tools. In the early 1700's, Western medicine adopted smallpox inoculation from Turkey, along with the idea of acquired immunity. The Germ Theory of disease had to replace spontaneous generation and miasma theory in the 1880's, however, before inoculation could successfully be applied to other diseases. Inquiry into acquired immunity led to the idea of the "antibody" in the 1890's, and the use of antiserum to identify bacteria. Immunostaining was invented in 1942 by repurposing antibody-dye conjugates originally intended as antibiotics. Monoclonal antibody-producing hybridomas were similarly invented in 1975 by repurposing techniques from virology and genetics.

A tale of the monoclonal anti-CD20 antibodies, in tribute to prof. Waclaw Szybalski (1921-2020).

Technical advances that lead to the era of targeted therapeutics demanded several milestones that were reached in the second half of the previous century. Professor Waclaw Szybalski was the first one to perform a stable gene transfer in eukaryotic cells. To do so, he used his own designed system consisting of HPRT-deficient cells and HAT selective medium. Moreover, the first-ever hybridoma cells were also constructed by Waclaw Szybalski's team. These spectacular achievements made him not only a forerunner of gene therapy, but also became a foundation for immunotherapy, as hybridoma and their selection by the HPRT-HAT system turned into a crucial technical step during production of monoclonal antibodies (mAbs). Herein, we present a story of anti-CD20 mAbs, one of the most successful lines of anticancer drugs. When looking back into history, the prototypic mAb rituximab was considered the biggest step forward in the therapy of B-cell malignancies. Nowadays, the second and third generations of anti-CD20 mAbs are approved in clinical use and numerous breakthrough studies on immune effector mechanisms were conducted with the aforementioned immunotherapeutics as a model.

Three decades of human monoclonal antibodies: past, present and future developments.

The hybridoma technique has been shown to be a most reproducible method for producing rodent monoclonal antibodies but poor results were obtained when it was used for generating human monoclonal antibodies. For immunotherapy, murine monoclonal antibodies are inadequate, whereas human monoclonal antibodies are virtually indispensable. Cellular, chemical, genetic and molecular methods to generate human monoclonal antibodies have been developed. Most often, the monoclonal antibodies for therapy are selected after deliberate vaccination, according to their high affinity towards an arbitrarily-chosen epitope of a pathogen or cellular antigen and therefore the selection is obviously skewed. A major hindrance of the production of therapeutic human monoclonal antibodies is the lack of an appropriate strategy to define and select the antibodies that would be effective in vivo. In contrast to antibodies induced by vaccination, there has been only a marginal interest in monoclonal antibodies which reflect antibodies of the innate immunity. In the future, human monoclonal antibodies that resemble antibodies that are ubiquitously present in sera of healthy individuals might serve as novel therapies in diseases such as Alzheimer's disease, where no other therapy exists.

[From the ancient serotherapy to naked antibodies: a century of successful targeted therapies]. / De la sérothérapie aux anticorps recombinants << nus >> : plus d'un siècle de succès en thérapie ciblée.

Monoclonal antibodies and molecular engineering have renewed the ancient serotherapy, multiplying the possibilities of therapeutic interventions and providing many new clinical successes! Standing back about this history allows us to better understand the evolution of concepts underlying the therapeutic use of antibodies, as well as the maturation of the tool itself. The different principles of therapeutic targeting will be successively tackled, from their sometimes hundred year-old conception until the most recent clinical developments: antibodies neutralizing toxins and soluble antigens, anti-microbial antibodies, cytotoxic antibodies, tumour-specific antibodies, cell function -modifying antibodies, etc. This overview will finally offer the opportunity to introduce a new pharmacological classification of the entire class of unconjugated -therapeutic antibodies.

Historical development of monoclonal antibody therapeutics.

Since the first publication by Kohler and Milstein on the production of mouse monoclonal antibodies (mAbs) by hybridoma technology, mAbs have had a profound impact on medicine by providing an almost limitless source of therapeutic and diagnostic reagents. Therapeutic use of mAbs has become a major part of treatments in various diseases including transplantation, oncology, autoimmune, cardiovascular, and infectious diseases. The limitation of murine mAbs due to immunogenicity was overcome by replacement of the murine sequences with their human counterpart leading to the development of chimeric, humanized, and human therapeutic antibodies. Remarkable progress has also been made following the development of the display technologies, enabling of engineering antibodies with modified properties such as molecular size, affinity, specificity, and valency. Moreover, antibody engineering technologies are constantly advancing to enable further tuning of the effector function and serum half life. Optimal delivery to the target tissue still remains to be addressed to avoid unwanted side effects as a result of systemic treatment while achieving meaningful therapeutic effect.

Herceptin.

The biology of the human epidermal growth factor (EGF) receptor-2 (HER2) has been reviewed numerous times and provides an excellent example for developing a targeted cancer therapeutic. Herceptin, the FDA-approved therapeutic monoclonal antibody against HER2, has been used to treat over 150,000 women with breast cancer. However, the developmental history of Herceptin, the key events within the program that created pivotal decision points, and the reasons why decisions were made to pursue the monoclonal antibody approach have never been adequately described. The history of Herceptin is reviewed in a way which allows the experience to be shared for the purposes of understanding the drug discovery and development process. It is the objective of this review to describe the pivotal events and explain why critical decisions were made that resulted in the first therapeutic to successfully target tyrosine kinases in cancer. New approaches and future prospects for therapeutics targeting the HER family are also discussed.

Tutorial on Monoclonal Antibody Pharmacokinetics and Its Considerations in Early Development.

The tutorial introduces the readers to the fundamentals of antibody pharmacokinetics (PK) in the context of drug development. Topics covered include an overview of antibody development, PK characteristics, and the application of antibody PK/pharmacodynamics (PD) in research and development decision-making. We also discuss the general considerations for planning a nonclinical PK program and describe the types of PK studies that should be performed during early development of monoclonal antibodies.

The story so far in R&D.

Human diseases are a significant cause of suffering and mortality and lead to a consequential need for effective therapies. The need for therapy is as old as human history itself. Therapy has progressed from an age of administration of herbal remedies and organ extracts to an era of serendipitous drug discovery, when the pharmaceutical industry was born, to the dominance of medicinal chemistry and more recently, to the revolutionary advances--genetic engineering and monoclonal antibody technology, high-speed technologies, gene therapy and the deciphering of the human genome--which promise the discovery of completely new targets for new medicines as well as the great potential of personalized therapy.

Natalizumab (Tysabri) treatment for relapsing multiple sclerosis.

BACKGROUND: Natalizumab (Tysabri), a humanized monoclonal antibody which binds to the alpha4beta1 integrins of leukocytes, blocks attachment to cerebral endothelial cells, thus reducing inflammation at the blood-brain barrier. Two pivotal randomized trials, 1 comparing active treatment to placebo and 1 comparing active treatment to placebo in relapsing multiple sclerosis (MS) patients receiving intramuscular interferon beta1alpha (Avonex), demonstrated significant efficacy for relapse control, decrease in sustained disability, and reduced numbers of new lesions on MRI. REVIEW SUMMARY: Because of safety issues raised by the appearance of 2 cases of progressive multifocal leukoencephalopathy in MS patients exposed to natalizumab for relapsing MS treatment has been restricted. This review briefly outlines the inflammatory nature of MS plagues, the mechanism of action of natalizumab and the pathogenesis of progressive multifocal leukoencephalopathy. The mandatory guidelines for natalizumab use in the treatment of MS with natalizumab are explained. Special concerns facing the therapist electing to prescribe natalizumab are mentioned. CONCLUSION: Natalizumab recently joined glatiramer acetate and beta interferon as an approved therapy for controlling relapsing MS. Unresolved safety issues currently restrict its use to monotherapy in patients who have had inadequate response to the other immunomodulating agents.

A selected history and future of immunoassay development and applications in clinical chemistry.

BACKGROUND: The first immunoassay was described by Berson and Yalow in 1959. Their work resulted in their receipt of the Nobel Prize in Medicine in 1977. Since this introduction, immunoassays have evolved considerably. METHODS: There have been several milestones that have led to the proliferation of modern immunoassays. The development of monoclonal antibodies from mouse hydridoma cells by Millstein and Kohler (Nobel Prize in 1984) enabled the production of high quantities of antibodies with well characterized epitope specificity. The first homogenous immunoassay (no separation step required) was the Enzyme Multiplied Immunoassay Technique (EMIT), which enabled adaptation of this assay onto automated chemistry platforms. EMIT was also one of the first immunoassay that made use of non-isotopic labels. Other non-isotopic labels became available such as chemiluminescence to improve the analytical sensitivity of immunoassays. The advantages of high-sensitivity immunoassays have created expanded diagnostic roles for some existing assays such as thyroid stimulating hormone for hyperthyroidism, C-reactive protein for cardiovascular risk assessment, and other applications. The development of instrumentation capable of automated heterogeneous immunoassays (separation step to improve sensitivity) has enabled movement of this technology from the "special chemistry" sections of a clinical laboratory into the "core" laboratory with other high-volume testing. CONCLUSION: Today, immunoassays play a prominent role in the analysis of many clinical laboratory analytes such as proteins, hormones, drugs, and nucleic acids. The future involves development of assays with higher sensitivities which will enable the discovery of new biomarkers for disease diagnosis, and technology that will enable simultaneous multimarker analysis of tests whose needs are naturally grouped together (e.g., cytokines and allergens).