Robert Huber was born February 20, 1937 in Munch, Germany. In 1956, he graduated from the Humanistische Karls-Gymnasium. He began studying chemistry at the Technische Hochschule (later Technische Universität) in Munch from which he received the Diploma in Chemistry in 1960. In 1963, he received a Dr. rer. nat researching the crystal structure of a diazo compound. He continued crystallographic studies at the University and the Physiologisch-Chemisches Institut der Universität München. In 1971, the University of Basel offered him a chair of structural biology at the Biozentrum and the Max-Planck-Gesellschaft the position of a director at the Max-Planck-Institut für Biochemie where he stayed until 2005. In 2005, he was made Director Emeritus. He also continued with the Technische Universität München, where he became Professor in 1976.

In 1967, he began crystallographic work on the insect protein erythrocruorin. The elucidation of this structure and its resemblance to the mammalian globins as determined by Perutz and Kendrew in their classical studies suggested for the first time a universal globin fold of the insect metamorphosis hormone ecdysone. In 1970, he started research on the pancreatic trypsin inhibitor, proteolytic enzymes and their natural inhibitors, proteases, their proenzymes, and complexes between them. He also studied the structure of immunoglobulin, the first glycoprotein to be analyzed in atomic detail. He elucidated the structure and the chemical nature of the selenium moiety in glutathione peroxidase and the structures of citrate synthase in different states of ligation and a very large multienzyme complex, heavy riboflavin synthase. Studies of proteins involved in excitation energy and electron transfer, lightharvesting proteins led to the structure of the reaction center.

For the determination of the three-dimensional structure of a photosynthetic reaction centre, he shared the Nobel Prize in Chemistry with Johann Deisenhofer and Hartmut Michel in 1988. He received the E. K. Frey Medal from the German Society for Surgery in 1972, the Otto Warburg Medal from the German Society for Biological Chemistry in 1977, the Emil von Behring Medal from the University of Marburg in 1982, the Keilin Medal from the London Biochemical Society and the Richard Kuhn Medal from the Society of German Chemists, 1987, and the Sir Hans Krebs Medal in 1992. He has also received numerous honorary doctorates and memberships in foreign chemical and biochemical societies.

He married Christa Essig in 1960 and they have four children.