Du Vigneaud graduated from Schurz High School in 1918. He began studying chemistry at the University of Illinois at Urbana-Champaign. He was influenced by lectures of Carl Shipp Marvel. After receiving his M.S. in 1924 he joined DuPont.
He married Zella Zon Ford on June 12, 1924. Restarting his academic career in 1925, he joined the group of John R. Murlin at the University of Rochester for his Ph.D thesis. He graduated in 1927 with his work, The Sulfur in Insulin.
After post-doctoral position with John Jacob Abel at Johns Hopkins University Medical School (1927-1928), he traveled to Europe as National Research Council Fellow in 1928-1929, where he worked with Max Bergmann at the Kaiser Wilhelm Institute for Leather Research in Dresden, and with George Barger at the University of Edinburgh Medical School. He then returned to the University of Illinois as a professor.
He joined Alpha Chi Sigma while at the University of Illinois in 1930.
In 1974 he suffered from a stroke which ended his academic career. One year after his wife's death in 1977, he died.