Goodpasture, the son of a lawyer, was born in Montgomery County, Tennessee. He was educated at Vanderbilt University, Nashville, and at Johns Hopkins University, where he gained his MD in 1912. After working as a pathologist for some years at Johns Hopkins and at Harvard, Goodpasture returned to Vanderbilt in 1924 as professor of pathology, a post he retained until his retirement in 1955.

In 1931 Goodpasture devised a method of virus culture that provided an enormous stimulation to virology. Before this, as viruses will grow only in living tissue, they could be studied experimentally either in a living host, or, after the work of Alexis Carrel in 1911, in vitro in a tissue culture. The first method was expensive and difficult to control while the second, before the advent of antibiotics, was susceptible to contamination by bacteria.

Goodpasture, in collaboration with Alice Woodruff, avoided such difficulties by providing a cheap living environment for viral growth – a fertile egg. Their first success was with fowl pox but within a year they had also grown both cowpox and coldsore viruses. Goodpasture went on in 1933 to show that attenuated cowpox vaccine could be produced in a purer and cheaper form in eggs than by the customary method of production in calf lymph.

Within a few years Goodpasture's technique had made possible the production of vaccines against yellow fever by Max Theiler and influenza by Thomas Francis. Thereafter eggs became as standard a part of the virologist's laboratory as the test tube.