Karl Otto Landsteiner (1868–1943). Physician–biochemist–immunologist

His childhood and early education

Landsteiner was born in Baden bei Wien near Vienna, Austria on 14 June 1868. His birth was just after the inception of the sciences of microbiology and immunology. ²³ His mother, Fanny Hess Landsteiner, came from a Jewish merchant family from Prossnitz in Moravia.^24–26 His father, Leopold Landsteiner, obtained a doctoral degree in law. But instead of practicing law, Leopold became a journalist and the co-founder of modern, liberal Austrian journalism. Before then, he had been a correspondent for German newspapers in Paris and been a lecturer in political science at the University of Lille. In 1848, the year of the Austrian Revolution, ²⁷ Leopold Landsteiner returned to Vienna to become with von August Zang the Editor-in-Chief of Die Presse, an important liberal daily newspaper in Vienna.

Leopold Landsteiner died in 1875 at age 58 when Karl was just seven years old. Karl had no siblings. Karl was placed under the guardianship of a family friend. But his mother principally raised him. Landsteiner attributed most of his success to the care and encouragement given to him by his mother. In fact, after her death in 1908, he kept her death mask on his wall.

Landsteiner was a good student and an excellent pianist. He attended the noted Wasa Gymnasium in the ninth district of Vienna. Afterwards, he attained an excellent education in universities not only in Austria, but also in Germany and Switzerland.

Landsteiner entered the University of Vienna Medical School in 1885 at age 17. That medical school was the second oldest one in the German-speaking world and was recognized for its high quality. At age 20, he served an obligatory year in the military. Landsteiner’s interest in medical science emerged during medical school when he began to do research in biochemistry. In that respect, the year after he graduated from the University of Vienna Medical School (1891), he published an article concerning the influence of diet of humans upon the ash composition of their blood. ²⁸

Landsteiner converted to Catholicism at age 20 and was christened Karl Otto Landsteiner at that time. A possible reason for the conversion was that Jews were not readily accepted in academic positions in Austria-Hungary in the late 19th century. In that respect, in some parts of Central and Western Europe where Jews were readily accepted, many of them decided to be assimilated into the general society. Conversion to Catholicism or another branch of Christianity therefore may have been a logical step for Landsteiner, because he apparently was already considering an academic career.

After graduating medical school, Landsteiner worked briefly with Dr. Otto Kahler (1849–1893), a professor of special pathology at the University of Vienna who described multiple myeloma and syringomyelia. From 1894 to 1895, he served in the First Surgical University Clinic in Vienna led by Eduard Albert (1841–1900), a pioneering orthopedist. But instead of seeking further clinical training, Landsteiner decided to expand his knowledge of human biochemistry, because he felt that field was where the future of medicine lay. He therefore spent five years in laboratories of four famous biochemists – Arthur Rudolf Hantzsch (1857–1935) and Roland Scholl (1865–1945) at Zürich (1891–1893) studying pyridine synthesis, ²⁹ Emil Fischer (1852–1919) at Würzburg, Germany (1893–1894) synthesizing the smallest possible molecule that contains both an aldehyde group and a hydroxyl group – glycolaldehyde, ³⁰ and Eugen Bamberger (1857–1932) at Munich, Germany (1896–1898) investigating phenylhydroxylamine. ³¹

In 1896, he became an assistant to the bacteriologist Max von Gruber (1853–1927) at The Hygiene Institute at Vienna. That seemed to bode well for Landsteiner, since von Gruber had discovered specific bacterial agglutination. However, working conditions at the Hygiene Institute in Vienna were poor. In 1896, he transferred to the Pathological-Anatomical Institute of the University of Vienna where he became an unpaid assistant to the pathologist/bacteriologist Anton Weischelbaum, who in 1887 was the first to isolate a causative agent (meningococcus) of cerebrospinal meningitis. ³²

Landsteiner conducted 3639 post-mortem examinations during his 15 years at The Pathological-Anatomical Institute. Despite this heavy workload, he investigated blood group antigens by using serological methods. The totality of his laboratory experiences allowed Landsteiner to become an expert with microbial and serological methods that he would use in much of his future research.

In 1903, Landsteiner became a Privatdozent at the University of Vienna. Then in 1908, Landsteiner was appointed as the director of the laboratories of the Royal-Imperial Wilhelmina Hospital in Vienna, where he remained until 1919. In 1911, he became Professor of Pathological Anatomy in the University of Vienna, but without a corresponding increase in his salary. He was, however, more interested in pathophysiology than anatomic pathology.

Discovers major blood group antigens in Vienna

Soon after he began working in Vienna, Landsteiner began investigating why some blood transfusions caused serious, even life-threatening reactions and others did not. But as Landsteiner emphasized in his 1930 Nobel Prize address, ⁶ there was a more fundamental scientific question that led to his research. “The problem raised by the discovery of biochemical specificities peculiar to a species – was to establish whether the differentiation extends beyond the species and whether the individuals within a species show similar though smaller differences.”

Since structural analyses of complex proteins were impossible in the early 20th century, Landsteiner turned to serological methods to investigate the basis of blood transfusion reactions. The approach was similar to one that Jules Bordet used to discover the interaction of specific antibodies to foreign red blood cells that led to agglutination and the lysis of those cells due to complement.^33,34

As Nevin C Hughes-Jones and Brigitte Gardner indicated in their 2002 historical review of red cell agglutination, ³⁵ Landsteiner was aware that when Adolf Crete (1847–1921) was a medical student in 1869 in Gottingen, he observed that blood brought in contact with serum from a different mammalian species agglutinated and hemolyzed. ³⁶ Although most of Crete’s experiments concerned intravascular lysis, in vitro mixing of red blood cells and serum from different animal species (for example, dog serum with rabbit red blood cells) also caused agglutination and hemolysis of the red blood cells.

In 1875, while Leonard Landois (1837–1902) was a professor and director of the Physiological Institute in the University of Greifswald in Germany, he published an extensive monograph on blood transfusions. ³⁷ Although the work was similar to Crete’s, Crete’s article was not referenced. However, both investigators concluded that a factor in serum interacting with the foreign red blood cells caused the agglutination and hemolysis of red blood cells.

The first suggestion of the existence of red cell antigens and serum agglutinins in what would be known as the ABO blood group system was found in a footnote in an article that Landsteiner published in 1900. ³⁸

Subsequently, Landsteiner demonstrated that when most specimens of human blood from most individuals were mixed with serum from other individuals, no agglutination occurred. ¹ But in some cases, agglutination occurred. Landsteiner found that the agglutination was as pronounced as when serum and red blood cells from different animal species were mixed. Further investigations demonstrated the agglutinations were not due to an illness in the serum donors.

After an extensive investigation, Landsteiner concluded that that there were four different types of human blood based upon the presence or absence of two red blood cell antigens (A and B). Both may be absent (type O), both may be present (type AB), or one may be present (type A or B). Furthermore, he discovered that the specific agglutinating antibodies were present only in those humans who lacked the specific blood type antigen to which the antibodies were directed.

The famous German immunologist, Paul Ehrlich, was opposed to Landsteiner’s interpretation of isoagglutinins. ³⁹ He asked what sense it made that “things are in the blood circulation directed against quite heterogeneous materials which under normal circumstances can never come into the picture” and “what good is it to the goat if it has in its blood something directed against the red cells or against spermatozoe of other animals”. Although Ehrlich was famous for his fundamental discoveries in immunology, ⁴⁰ Landsteiner’s findings were correct. Almost immediately, Landsteiner’s discoveries of the major blood groups led to safe blood transfusions. Of longer term significance is that his discovery was the first demonstration of an inheritance pattern of human proteins. ⁴¹ Indeed, red cell typing would later be used to determine paternity.

Experiments with syphilis

In 1906, Landsteiner and Ernest Finger (1856–939), the Chief of the Clinic for Venereal Diseases and Dermatology in Vienna, demonstrated that the cause of syphilis, Spirochaeta pallida, could be experimentally transmitted to Old World monkeys. ²⁰ At the same time, Landsteiner and Viktor Mucha, a scientist from the Chemical Institute at Finger's clinic, showed that dark-field microscopy could be used to identify Spirochaeta pallida. ¹⁹

August Paul von Wassermann (1866–1925), a German bacteriologist, together with Albert Neisser (1855–1916), a German dermatologist and venereologist, and Carl Bruck (1879–1944), a German dermatologist and venereologist, developed the first serologic test for the diagnosis of syphilis. ⁴² Landsteiner identified that the antigen involved in the Wassermann reaction was a lipoid substance. ⁴³ The antigen was later found by others to be diphosphatidylglycerol.

Discovers the pancreatic pathology in cystic fibrosis

During the time that Landsteiner was investigating the human blood group antigens, he was doing virtually all of the autopsies at the Pathological-Anatomy Institute in Vienna. In 1908, he chanced upon a newborn female who died on the fifth day of life from intestinal obstruction due to meconium ileus. ⁹ The histopathological changes in the pancreas were as follows.

The pancreas shows a very increased amount of inter- and intra-lobular connective tissue. This connective tissue has coarse fibers. In the areas of the lobules nuclei are rich and in some places a rather large amount of an infiltrate with small round cells. The infiltrates are in part circumscript with round forms and in part stretched into larger areas containing interacinar tissue. While the islets show no changes, the secreting parenchyma are affected. It appears that the mass of secreting tissue is reduced. The cells are small and flat. They are frequently transparent, and in these cells, secretion is perceived to be impeded in the form of homogenous discharge or several round flat cells are found next to one another. The discharge channels are more or less scattered to a high degree and contain homogenous, solid, or granular secretions. Here and there are radial, eosin colored masses. The appearance of these secretion blockages is not equally distributed, but rather affect individual acini in higher measure than others. However, the process is found in all parts of the organ.

Experimentally transmits poliovirus in Vienna and Paris

Because an epidemic of paralytic poliomyelitis was raging in Austria and nearby countries in 1908, the leaders of the hospital at Universität von Wien asked Landsteiner to try to determine the cause of the disease. They hoped that if a cause could be found, then a vaccine against the agent could be developed. The hope was that such a vaccine could be based upon the vaccines that Pasteur developed against chicken cholera, ⁴⁵ sheep anthrax, ⁴⁶ and human rabies. ⁴⁷

Although others had reported that bacteria were cultured from spinal cords or cerebrospinal fluid from cases of paralytic poliomyelitis, Landsteiner discounted those findings since no bacteria were seen in stained affected tissues by light microscopy. His interpretation was that the bacteria were contaminants, rather than pathogens. He thus predicted that poliomyelitis could be due to a microbial agent smaller than bacteria. But first he had to establish whether the infection could be transmitted to experimental animals.

Landsteiner used spinal cord tissues from a nine-year-old boy, who died in November 1908 from paralytic poliomyelitis. No bacteria were detected in the child’s spinal fluid or spinal cord by direct microscopy or in vitro cultures. Landsteiner tried unsuccessfully to transmit human poliomyelitis into mice, rabbits, and guinea pigs by injecting them with a preparation of affected spinal cord tissue. Landsteiner then tried to infect two Old World monkeys (a Cynocephalus and a Macaca rhesus) that are more closely related to humans than those other species. A bacteriologically sterile suspension of the child’s infected spinal cord was injected intraperitoneally into both animals. The Cynocephalus hamadryas became ill six days after the injection and died two days later. The histological findings were consistent with bulbospinal poliomyelitis. The Macaca rhesus lived but developed complete flaccid paralysis of the legs 12 days after the injection. The anterior horn motor neurons in the spinal cord of the Macaca were damaged or destroyed. The transmissible agent in either animal was not detected by light microscopy or by in vitro cultures. ⁸

Landsteiner wished to test whether the infectious agent that caused paralytic poliomyelitis was smaller than bacteria or fungi. Filtration experiments were required to test that possibility, but no other Old World monkeys were available for those experiments in Vienna. Landsteiner therefore asked the director of l’Institut de Pasteur de Paris, Emil Roux, whether a researcher at l’Institut would be interested in conducting those experiments with him and whether Old World monkeys were available for those studies. Constantin Levaditi (1874–1953), a Romanian physician and microbiologist, agreed to help him conduct the experiments. That was fortunate because Levaditi was an excellent scientist who later became famous in his own right ⁴⁸ Experiments carried out at l’Institut showed that the poliovirus was filterable, whereas bacteria and fungi were not.^49,50 Landsteiner and Levaditi concluded that the transmissible agent was a virus. Simon Flexner and his colleague Paul Lewis at the Rockefeller Institute at New York City quickly confirmed the findings.^51,52

Because Landsteiner’s mother had died in 1909, Landsteiner requested a leave of absence to spend more time at l’Institut de Pasteur de Paris to continue to investigate polioviruses with Levaditi. The directors at University of Vienna granted him a leave of absence. During that period, Landsteiner and Levaditi determined that polioviruses could be detected in the pharynx, tonsils, salivary glands, and lymph nodes as well as the spinal cord^53,54 and in the amygdala and nasal secretions of infected non-human primates. ⁵⁵ That laid the foundation for further understanding the biology of polioviruses. ⁵⁶ He and Levaditi also found that the virus could be neutralized in the laboratory when mixed with the serum of a convalescing patient. It was the first demonstration of neutralizing antibodies against polioviruses.

Landsteiner and Levaditi experimented with ways of chemically deactivating polioviruses in vitro in hopes of creating a safe, effective poliovirus vaccine. ⁵⁷ They later attempted to create an attenuated poliovirus vaccine by treating virus-infected tissue at 56℃ for 30 to 60 min. However, the work was abandoned because the inoculated monkeys developed paralytic poliomyelitis.

Landsteiner did not conduct further research concerning polioviruses for a number of reasons. When he returned to Vienna in 1911 where he was promoted to the position of Professor Extraordinarius, there were no further funds for the purchase and upkeep of experimental Old World monkeys in Vienna. Indeed, a method for the in vitro cultivation of poliovirus that led to a further elucidation of the nature of the virus would not be developed until 1949, ⁵⁸ and the first safe successful poliovirus vaccine would not be produced until 1954. ⁵⁹

During the First World War, Landsteiner served as a physician in an Austrian military hospital caring for the many causalities of the war. ⁶⁰ During the war, in 1916, at age 48, he married Leopoldine Helene Wlasto. Their only child, Ernst Karl, was born the following year.

Discovers haptens

In 1919, German Austria was dissolved by the Treaty of Saint Germain, which ceded German-populated regions in Sudetenland to Czechoslovakia, German-populated Tyrol to Italy and a portion of the south to the Kingdom of Serbs, Croats, and Slovenes. It also barred union with Germany unless the League of Nations consented.

The new republic of Austria was in dire economic straits. Therefore, it was impossible for Landsteiner to continue his research. In 1919, he accepted a post as prosector in the small Catholic Ziekenhuis Hospital in The Hague in the Netherlands. To improve his financial situation, he also worked in a small factory producing old tuberculin for therapeutic purposes advocated by the discover of Mycobacterium tuberculosis, Robert Koch.^61,62

Before he left Vienna in 1917, he discovered haptens – small molecules such as polysaccharides or lipids that combine with specific antibodies but cannot elicit an immune response unless they are attached to a large carrier such as a protein.^10,11 Despite a heavy workload at the Catholic Ziekenhuis Hospital, he continued to discover new haptens.^12–14

Research at the Rockefeller Institute in New York

The financial situation of the Catholic Ziekenhuis Hospital in the Hague was as insecure as Landsteiner had encountered in Vienna. As fate would have it, when Simon Flexner, the head of the Rockefeller Institute of Medical Research in New York and the co-discoverer of the experimental transmission of polioviruses, learned of Landsteiner’s plight, he offered him a research position in his institute. Landsteiner immediately accepted and moved with his wife and son to New York that year, 1923, at age 55.

At the Rockefeller Institute, Landsteiner constantly sought funding for his research. He had a small laboratory and few supplies until his Nobel Prize award brought a larger laboratory, supplies, and more staff. His main interest at this time was in the chemical and immunological basis of skin sensitization and allergy. Together with Merrill W Chase, he showed that skin sensitivity to certain chemicals was induced by allergens formed by combinations of these components (haptens) with host proteins. Experiments in which lymphocytes from a sensitized animal were transferred into a normal recipient were the basis of further studies on the cellular aspects of contact dermatitis and other cell-mediated immune responses. ⁶³

At the Rockefeller Institute, Landsteiner conducted further research concerning blood group antigens and other important subjects until his death in 1943 at age 75. He and his colleague Philip Levine enlarged the knowledge of human blood group antigens by discovering blood group antigens M, N, and P. ⁶

In 1940, Landsteiner and Alexander S Wiener reported their discovery of the Rhesus (Rh) blood type in human blood. ⁶⁴ The discovery was made by immunizing rabbits with red blood cells from a Rhesus macaque. ⁶⁵ Rabbit antibodies to the Rhesus red blood cell antigen also agglutinated and hemolyzed human red blood cells from certain individuals. The antigen that induced this immunization was designated as Rh factor to indicate that rhesus blood had been used to produce the serum.

In addition to his pioneering studies concerning immunohematology, Landsteiner and Clara Nigg were the first to culture Rickettsia prowazeki in living tissue.^21,22

Landsteiner retired from the Rockefeller Institute at age 71 in 1939 but he continued to work until his death.

One wonders why Landsteiner did not attempt to develop a vaccine against polioviruses after he came to the Rockefeller Institute for Medical Research. He and Simon Flexner, the Founder and past Director of the Institute, were the first to experimentally transmit the virus to Old World monkeys. Both had independently tried to create a poliovirus vaccine earlier, but had failed. It is unknown whether Landsteiner considered developing such a vaccine in the 1920s and early 1930s. But two tragic experiments in children in the 1930s may have dissuaded him from reexamining the possibility.

In 1934, Dr. John Albert Kolmer, Professor of Medicine and Director of the Research Institute of Cutaneous Medicine at Temple University, developed a poliovirus vaccine from infected monkey brain tissues treated with the emulsifying agent sodium ricinoleate. ⁶⁶ Kolmer chose ricinoleate because it was believed that it was a detoxifying agent. He felt that the treatment would produce an attenuated poliovirus vaccine that would be more immunogenic than a killed viral vaccine. Kolmer was incorrect. After he injected the experimental vaccine into brains of non-infected monkeys, the animals seemed well and did not develop paralytic poliomyelitis when they were subsequently injected with live polioviruses. Consequently, in 1935, Kolmer carried out a large-scale trial of the vaccine in children. Tragically, many vaccinated children developed paralytic poliomyelitis. Nine died, probably of bulbospinal or bulbar poliomyelitis. ⁵⁶ In retrospect, the vaccines must have contained live polioviruses.

In 1934, Dr. Maurice Brodie and Dr. William H Park at New York University produced a formaldehyde-killed poliomyelitis vaccine from ground-up spinal cords obtained from monkeys infected with polioviruses. Formaldehyde was chosen because it was a widely used tissue fixative. The tissue preparation produced specific antibodies to polioviruses when injected into uninfected monkeys. They then injected the experimental poliovirus vaccine into about 75 children. ⁶⁷ Serum antibodies to the virus were formed. No untoward effects occurred.

Consequently, Brodie and Park organized an extensive trial of this experimental poliovirus vaccine. In 1935, three thousand children received the experimental vaccine. Unfortunately, the vaccine was not protective, many injected children became ill, and one died. ⁵⁶ The nature of their illnesses following the administration of the vaccine was never revealed, but it may have been an autoimmune encephalopathy due to systemic exposures to antigens from monkey spinal cords that elicited antibodies that cross-reacted with human brain antigens.

Those two disastrous trials brought research into poliovirus vaccines to a halt. It is somewhat ironic that the first successful poliovirus vaccine was patterned after Brodie and Park’s, and that the developer of that vaccine, Jonas Salk, was a medical student at New York University when Brodie and Park were experimenting with their poliovirus vaccine.

Importance of Landsteiner’s discoveries

Landsteiner’s discoveries were of great medical importance. (1) His demonstrations of the major blood group antigens (types A and B) and their isoagglutinins were the basis for developing safe human blood transfusions that have saved millions of lives. Other researchers later discovered that the two major blood group antigens were glycoproteins and glycolipids. However, the exact biological functions of those antigens were never determined. Furthermore, the origin of antibodies to group A and B antigens remains uncertain, although it is likely that they arise from exposures to food, plant, or microbial antigens that cross-react with A and B glycoprotein antigens. ⁶⁸ Moreover, Landsteiner was the first to discover genetic markers responsible for the synthesis of certain proteins – the major blood group antigens. ⁴¹ (2) His elucidation of other blood group antigens (M, N, and P) provided a way to conduct forensic investigations and paternity tests long before genetic determinants came into use. (3) The demonstration of the Rh antigen led to an understanding of the cause of most cases of hemolytic disease of the newborn and of the methods used to treat or prevent those adverse immunological reactions by exchange transfusions and antenatal Rh immunization, respectively. (4) His discovery of haptens led to a further elucidation of the requirements for the formation of specific antibodies. Indeed, Landsteiner was proudest of his work on antigen specificity that laid the foundation for the field of immunochemistry. (5) His were among the first investigations of hypersensitivity reactions to simple chemical agents. (6) He also experimentally demonstrated that certain foreign proteins produced anaphylaxis only after they were partially digested by pepsin. ⁶⁹ Whether that occurs in the genesis of certain allergic reactions to certain food antigens remains to be investigated. (7) His demonstration that poliomyelitis was due to a transmissible virus culminated in the development of poliovirus vaccines in the 1950s that brought the epidemics of poliomyelitis to a close. (8) Landsteiner with Merrill W Chase demonstrated that cutaneous sensitivity to simple chemicals could be transferred to non-sensitized guinea pigs and that the sensitization was due to lymphocytes and not to antibodies. ⁶³ At first Landsteiner did not believe the results. However, the finding was the precursor to Chase’s discovery of cellular immunity that began with Ilya Metchnikoff’s recognition of macrophages in the late 19th century.^40,70

One remarkable late achievement was his book “The Specificity of Serological Reactions”, the third edition of which was published after his death in 1945. ¹³ The book at that time was the most extensive survey of the structures and types of antigens, the serological and chemical basis of the nature and creation of antibodies, and their reactions with antigens. It was the primer for the molecular revolution in immunology that began after Second World War and continues to this day. The book is also important because it contains a complete list of Landsteiner’s 364 publications.

In his book, Landsteiner carefully reviewed the question of how specific antibodies were formed. Although Paul Ehrlich correctly imagined that preformed antibodies on cells would be receptors for antigens and that the complexing of cell-bound antibodies to specific antigens would stimulate the cells to produce and secrete more antibodies against the specific antigen, ⁷¹ it was difficult for Landsteiner or other immunologists to accept it. In fact, Ehrlich’s side chain theory of antibody receptors was not proven to be membrane immunoglobulin antigen receptor on B cells until the 1970s, ⁷² and membrane-bound antigen receptors on T cells were not demonstrated until 1980s. ⁷³

During Landsteiner’s time, most immunologists favored a template mechanism whereby antigens impressed themselves upon the structure of protein molecules that were to become antibodies. Landsteiner was not convinced because the numbers of antibody molecules that were formed exceeded the much smaller numbers of eliciting antigen molecules. Although the template theory of specific antibody formation proved to be incorrect, it set the stage for the investigations that led to the realization that specific antibodies formed from a complex mechanism that involved the rearrangement and recombination of specific variable, joining, and diversity genes. ⁷⁴

Landsteiner lived before many of the laboratory methods that allowed scientists to investigate in depth the complexities of the immune system including the physical chemistry of the myriads of its protein components. The methods included electrophoresis, immunofluorescence, monoclonal antibodies, flow cytometry, amino acid sequencing, and molecular genomics that would revolutionize the field.

During his time in the United States, Landsteiner was very much respected by other immunologists. In that respect, he was elected as President of the American Association of Immunology in 1927, served as a councilor to that organization a few years before and after his presidency, and was on the Board of Editors of the Journal of Immunology from 1937 to 1942 and the Associate Editor in 1943. He was also elected to the American Philosophical Society in 1935 for his accomplishments that furthered medical science and the health of many thousands of people.

In addition to the Nobel Prize, Landsteiner was awarded the Hans Aronson Foundation Prize in 1926 for his accomplishments in microbiology and immunology, the Paul Ehrlich Medal in 1930 for his accomplishments in immunology, the Dutch Red Cross Medal in 1933 for the important deeds he performed, and the Albert Lasker Clinical Medical Research Award in 1946 for his research that saved or improved the lives of many millions of people.

He remained dedicated to science until the end of his life. In that respect, his fatal illness, a massive heart attack, was first manifest when he was working at the laboratory bench.

In conclusion, Karl Landsteiner was modest, self-critical, and dedicated to advancing medical science. We suspect the rigor and structure of chemistry gave his thinking a scaffold that helped to keep him on the right track through his many different types of research. Landsteiner did research without the aid of elaborate equipment required for research in the 21st century. But that lack was compensated for by Landsteiner’s clarity of thought, powers of detective and inductive reasoning and knowledge of biochemistry. The scope of his achievements has few equals for his time or thereafter.