Sanger determined the complete
amino acid sequence of
insulin in 1955. In doing so, he proved that
proteins have definite structures. He began by degrading insulin into short fragments by mixing the
trypsin enzyme (that hydrolyses the peptide/amide bonds between amino acids that make up the primary structure of proteins) with an insulin
solution. He then undertook a form of
chromatography on the mixture by applying a small sample of the mixture to one end of a sheet of filter paper. He passed a
solvent through the filter paper in one direction, and passed an
electric current through the paper in the opposite direction. Depending on their
solubility and
charge, the different fragments of insulin moved to different positions on the paper, creating a distinct pattern. Sanger called these patterns “fingerprints”. Like human
fingerprints, these patterns were characteristic for each protein, and reproducible. He reassembled the short fragments into longer
sequences to deduce the complete structure of insulin. Sanger concluded that the protein insulin had a precise amino acid sequence. It was this achievement that earned him his first
Nobel prize in Chemistry in 1958.
In 1975, he developed the
chain termination method of DNA sequencing, also known as the
Dideoxy termination method or the
Sanger method. Two years later he used his technique to successfully sequence the genome of the
Phage Φ-X174; the first fully sequenced DNA-based genome. He did this entirely by hand. This has been of key importance in such projects as the
Human Genome Project and earned him his second
Nobel prize in Chemistry in 1980, together with
Walter Gilbert. The only other laureates to have done so were
Marie Curie,
Linus Pauling and
John Bardeen. He is the only person to receive both prizes in chemistry. In 1979, he was awarded the
Louisa Gross Horwitz Prize from
Columbia University together with
Walter Gilbert and
Paul Berg, co-winners of the 1980 Nobel Prize in Chemistry.
Sanger also produced a revolutionary way of transcribing DNA using ethanol as a buffer.