It is not an exaggeration to describe César Milstein's contribution to science and medicine as the most important immunological advance of the century. His discovery of the method to produce monoclonal antibodies reinvented the field of immunology. The ability to make monoclonal antibodies at will in the test tube and in unlimited quantities, to any sort of antigen - whether an interesting chemical, infectious micro organism, cancer or normal cells - opened numerous new and unforeseen avenues for research, many with medical implications. It was therefore not surprising to learn that César got the largest number of nominations from the distinguished panel of scientists for the Nobel prize that he shared in 1984 with Georges Köhler and Nils Jerne.
However, the climb to the top was not an easy one. César was born on October 8 19 in a provincial Argentina town as the second son to his school teacher mother and his father, who had emigrated alone to Argentina from the Ukraine at the age of 14. His parents saw to it that their three sons got a university education. César studied chemistry at Buenos Aires and did a PhD on the active site in enzymes. In 1958, he won a British Council research scholarship to the biochemistry department of Cambridge and worked with Fred Sanger (who was awarded his first Nobel prize in December that same year). Having collected a second PhD, he returned to Buenos Aires in the hope of contributing to Argentine science. However, because of political interference in the laboratory, he returned to Cambridge at the invitation of Sanger who by that time had joined Max Perutz in the newly formed MRC Laboratory of Molecular Biology.
After his return to Cambridge César started to work on antibodies and within a few years he became an internationally acknowleged leader in the field. In 1966 he published a Nature paper with Sydney Brenner about the origin of antibody variation proposing that this resulted from mutations in the genes of antibodies. He was particularly interested in the genetic mechanisms by which humans or animals produce millions of different antibodies (from which the immune system selects those that bind to antigen), and also in the mechanisms by which the binding affinity of antibodies is improved on re-exposure to antigen.
One of the ways he chose to tackle this question was by using murine myeloma cells which are the malignant proliferation of the antibody-producing white blood cells. At that stage there were no tissue culture facilities in the MRC building, and César had no experience in this work. He was allocated a small, windowless basement store room as his tissue culture suite.
When I joined Cambridge University's department of haematology in 1969 (which was located in the MRC building) I was given a spacious virus and tissue culture laboratory to study and grow cells from leukaemia patients. Since my laboratory was next door to César's we met shortly after my arrival and on a daily basis for years to come. By that time I had already considerable experience with in vitro cell culture and since César was a beginner I became his "cell doctor".
By this time Richard Cotton joined César on a postdoctoral fellowship. They were attempting to fuse the mouse myeloma cells with the rat myeloma cells in order to study the antibody production by the hybrid cells. At that time the only agent that was known to fuse cells was inactivated Sendai virus. Since I produced it for my own experimental studies I offered to produce it for Cesar's studies as well. In 1973 César and Cotton published a Nature paper about the "Fusion of Two Immunoglobulin Producing Myeloma Cells". In fact it was the first antibody producing hybridoma but it was created by two malignant cells that readily grew in the laboratory and the specificity of the antibodies it secreted was not known.
Following this study, Cotton decided to return to Australia and César had to look for a new postdoctoral researcher to take forward the cell fusion project. Georges Kohler had just completed his PhD at the Basel Institut of Immunology and was attracted to the project. He joined César's group and also started to learn cell culture with my help.
Within two years Kohler and Milstein had succeeded in fusing the mouse myeloma with mouse lymphocytes that had been taken from an immunised animal immunized with sheep red blood cells. They were able to identify and grow hybrid cells producing a single species of antibody (a monoclonal antibody) against the sheep red blood cells. In their publication they pointed out that the method was rather general and should be useful to produce monoclonal antibodies against any antigen, and also that this had some important medical implications.
Although César's greatest talent was in innovative basic research he also realised the commercial potential of monoclonal antibodies and therefore he suggested in a letter to MRC head office that his laboratory's invention should be patented. Unfortunately, it appears that the National Research and Development Corporation (NRDC) which had a monopoly over MRC inventions did not think César's idea about monoclonal antibodies was worth patenting. It took a narrow view and said that it would have been much more interesting if César produced antibodies other than those against sheep. As a result the potentially enormous royalty income on the key step of what became a multibillion pound industry was lost. When Margaret Thatcher became prime minister and learned about the failure to apply for a patent it is said (somewhat unfairly) blamed Sir James Gowen who was the head of the MRC at the time.
Within a few years a very large number of laboratories around the world had started to produce monoclonal antibodies to an ever increasing number of targets. The availability of monoclonal antibodies revolutionised both basic and clinical research. For example, it enabled the first purification of interferon which was done in César's laboratory by David Secher. Monoclonal antibodies were used to enable an accurate diagnosis of different leukaemias and are used routinely in every medical centre to measure the levels of many blood proteins. When the antibodies are labelled with radioactive chemicals they can be used to detect and localise cancer tissue while an increasing number of antibodies are used as 'magic bullets' to destroy malignant tissue in cancer patients.
The 1970s was a golden era for César's research and for those who followed in his footsteps. In 1972 he published a Nature paper about the molecular mechanism and the process involved in the secretion of antibodies. This research topic was subsequently followed by Günter Blobel who two years ago received a Nobel prize. Due to his inspiration and support, two other MRC scientists, Greg Winter and Michael Neuberger, made further ground-breaking discoveries in the development of new methods to produce humanised and finally human monoclonal antibodies. As a result Cambridge can claim all the important discoveries and development in the monoclonal antibody field.
In spite of his official retirement in 1995, César continued to be an active researcher. In July 2000, the MRC organised a conference to celebrate the 25th anniversary of his discovery of the method to produce monoclonal antibodies. Not only scientists took part in this conference but also politicians who knew of his important contribution to science.
César's interests were not confined to research. He followed the world news on a daily basis and held opinions on important events. He was interested in numerous facets of the arts (often going to the theatre and concerts in London) and was an excellent cook. César was also a man with great sensitivity to human rights and justice. He was an advocate for peace and compromise in national conflicts and was proud of his family's Jewish heritage. In spite of the large number of international prizes he won, César remained the same modest and intellectually generous person that I first met in 1969. He remembered my early help with the cell culture and as a supplier of Sendai virus and invited me to take part in the Nobel celebrations in Stockholm.
His mother died at the age of 70 from a heart attack. Over 25 years ago César discovered that he had probably inherited his mother's vascular genes and so resorted to a strict diet and endless walks. When more serious cardiac problems developed, he was looked after by the excellent cardiologists at Papworth Hospital, one of whom told him that he felt that his major contribution to science was keeping César alive. César reciprocated their admiration and was full of praise for his doctors' efforts.
For nearly 50 years he was very happily married to Celia, a research scientist for many years - she was the one who saw to it that he never missed his doctor's appointments. Cesar's death has robbed us of one of the best-loved and most important scientists of the past century.
Abraham Karpas
Assistant director of research
Department of haematology
University of Cambridge
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