Lehninger was born in Bridgeport, Connecticut, U.S.. He received his Bachelor of Arts from Wesleyan University (1939) and went on to pursue both his Masters of Science (1940) and Ph.D. (1942) at the University of Wisconsin—Madison.

After receiving his doctorate in biochemistry, he held various faculty positions at the University of Wisconsin—Madison and the University of Chicago. In 1952, Lehninger came to the Johns Hopkins University School of Medicine assuming the title of DeLamar Professor of the Department of Biological Chemistry. He served in this position until 1978, when he was appointed to the role of University Professor of Medical Sciences. This position was created specifically for Lehninger, to honour his service to Johns Hopkins University, his pioneering scientific efforts and his dedication to his students. He held this title, and proudly served the scientific community, until his death in 1986. Posthumously, his memory is honoured through The Albert Lester Lehninger Memorial Lectureship at Johns Hopkins University.

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