Classic Articles:
O'Farrell 250 (10): 4007
J. Biol. Chem., Vol. 281, Issue 32, 26, August 11, 2006
Classics
The Development of Two-dimensional Electrophoresis by Patrick H. O'Farrell
Nicole Kresge,
Robert D. Simoni, and
Robert L. Hill
High Resolution Two-dimensional Electrophoresis of Proteins
(O'Farrell, P. H. (1975) J. Biol. Chem. 250, 4007–4021)
Patrick H. O'Farrell received his B.Sc. in 1969 from McGill University in Montreal, Quebec. He then went on to graduate school at the University of Colorado, Boulder, where he worked with Jacques Pène. At Boulder, O'Farrell isolated a series of mutations affecting development in the colonial algae, Volvox. His goal was to define the normal pattern of gene expression during development and the effects that the mutations had on this regulatory program. Similar studies had been done in bacteriophage using sodium dodecyl sulfate gel electrophoresis to separate, identify, and quantify the proteins. However, Volvox has approximately 100 times the genetic complexity of bacteriophage and thus needed a separation system with a much higher resolution. Although an adequate system did not exist, two-dimensional methods for increased resolution in chromatography and electrophoresis had been well established. So, O'Farrell recalls, "I set out to combine the two most powerful electrophoresis methods available" (1). This is the subject of the Journal of Biological Chemistry (JBC) Classic reprinted here.
O'Farrell realized that to optimize resolution, he would need to separate the proteins according to independent parameters. He decided to use isoelectric focusing in the first dimension and sodium dodecyl sulfate electrophoresis in the second dimension. This permitted the simultaneous determination of molecular weight and isoelectric point for the proteins. Because the two parameters are unrelated, it was possible to obtain an almost uniform distribution of protein spots across the two-dimensional gel. Using his technique, O'Farrell was able to resolve 1100 different components from Escherichia coli and predicted his system should be capable of resolving up to 5000 proteins.
O'Farrell's advisor, Pène, left Boulder around this time, but O'Farrell stayed on to complete his degree under the sponsorship of David Hirsh. After defending his thesis, O'Farrell submitted his two-dimensional electrophoresis manuscript to the JBC and moved to San Francisco to do postdoctoral studies with Gordon Tomkins in the Department of Biochemistry and Biophysics at the University of California, San Francisco (UCSF). Several months later, O'Farrell received a letter from the JBC rejecting his paper on the basis of two unfavorable reviews. The reviewers had concluded that the manuscript appeared to be "highly speculative in places and to be extrapolated in terms of usefulness far beyond what the author has any reason to expect." Fortunately, with the help of several members of the journal's editorial board, the initial decision was reversed, and the paper was published. O'Farrell's JBC Classic went on to be one of the 50 most highly cited items in Web of Science. As of January 2004, it has been cited in over 16,244 publications.
In 1975 Tomkins passed away, and O'Farrell moved to Bruce Alberts' laboratory at UCSF. He remained with Alberts until 1979 when he was appointed Assistant Professor in the Department of Biochemistry and Biophysics. O'Farrell was promoted to Associate Professor in 1985. Currently, he is a professor in the biochemistry and biophysics department at UCSF. He is also a member of the UCSF Biomedical Sciences Program and Herbert Boyer Program in Biological Sciences as well as a member of the UCSF Comprehensive Cancer Center. O'Farrell has received several honors and awards, including the Sarstedt Research Prize. He is a Fellow of the Jane Coffin Childs Memorial Fund for Medical Research and a Senior Fellow of the California Division of the American Cancer Society.
REFERENCES
- O'Farrell, P. H. (1982) High-resolution two-dimensional electrophoresis of proteins. Curr. Contents 51, 18
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
Copyright © 2006 by the American Society for Biochemistry and Molecular Biology.
