Along with his preliminary results, Itano suggested in a 1948 report that there were eight other options for immediate research,
one of which eventually proved successful: "electrophoretic studies on normal and sickle cell hemoglobin." In his synopsis
about electrophoresis, Itano briefly mentioned the preliminary status of electrophoretic studies on hemoglobin and its possible
utility for his research. Itano stated later that he initially tried electrophoresis because the apparatus was available and
required small amounts of blood. At the time, Caltech had a problem getting adequate samples of blood from people with sickle
cell anemia. Pauling finally obtained a regular supply in May 1949 from Dr. George Burch of Tulane University.
Electrophoresis uses electric currents to examine substances that have varying surface charges, and therefore is especially
helpful in analyzing proteins. Protein samples in a buffer solution are run through the electrophoresis apparatus, which creates
a moving boundary. Moving boundary electrophoresis indicates whether a protein has an overall positive or negative charge
on its surface area. The rate of a moving boundary can be manipulated by changing the pH of the solution and hence, the charge
of the particles.
Since moving boundary electrophoresis was a relatively new technique in 1948, only about fifty laboratories in the United
States had electrophoresis machines. Mass production of the apparatus began after 1950, so that prior to mid-century, laboratories
built them. Pauling asked the Rockefeller Foundation in 1941 for money for immunology research, including the construction
of a Tiselius electrophoresis apparatus. Caltech received enough money from the Rockefeller Foundation to build the machine,
to which Stanley M. Swingle, a general chemistry instructor at Caltech, made improvements. It was this machine that Itano and Seymour Jonathan Singer used for the experiments reported in the 1949 article.
Pauling's request for money to build an electrophoresis machine was most likely influenced by Karl Landsteiner's infatuation
with the device. Landsteiner, Pauling's mentor in immunology, analyzed blood sera with electrophoresis beginning in 1908.
When Landsteiner learned of Arne Tiselius's improved apparatus in 1937 he immediately began using the technique as a standard procedure in his researches.
Click images to enlarge
Cartoon caricatures of Linus Pauling, Jean Timmermans, Arne Tiselius, L. H. Lampitt, Robert Bienaime, B. C. T. Jansen, Viscount
Leverhulme, Wallace Akers, Marston T. Bogert, Robert Robinson, Paul Karrer and Professor Vesely. June 18, 1947.
Letter from Stanley Swingle to Linus Pauling. May 2, 1948.
"The item of $7,500 for apparatus, supplies, animals would permit us to use the large number of animals required for some
of our projected researches, and should permit also the construction of a Tiselius apparatus for the electrophoretic separation
of antibody fractions by the suggested method of combination with charged haptens, and for other investigations."
January 2, 1941