January 2012

W. Neal Burnette: the man behind the Western blot

Going west
At the end of 1977, Burnette took on another postdoctoral fellowship with Robert Nowinski at the Fred Hutchinson Cancer Research Center in Seattle. Nowinski, who went on to become the founder of the biotechnology company ContraFect, describes Burnette as an “eager and enthusiastic” postdoctoral fellow who worked fairly independently.

Nowinski’s group was prominent in the retrovirology field and was engrossed in the analysis of antigenic epitopes of retroviral structural proteins. But they were using tedious chromatographic separations and radioimmunoassays to probe each individual protein of the viral capsid with a series of antibodies. Burnette offered to find a way to speed up the process so that all the proteins in a viral capsid could be tested with an antibody in a single shot. “When you ask a carpenter to do something, the tool he’s always going to use is a hammer,” says Burnette. “My tools were SDS-polyacrylamide gels and immunoassays.”

The main development of the technique took Burnette two weeks in 1979 followed by a few more weeks of tweaking. One of the major problems Burnette had to grapple with was how to get antibodies to bind to proteins that were separated in the polymer matrix of a gel. But in a moment of inspiration, he realized that, just like the DNA and RNA blots that were all the rage at the time, he could make a replica of the gel-resolved proteins and use the replica for the immunoassay.

DNA blotting was called Southern blotting after its inventor, Edwin Southern at Oxford University (2). RNA blotting, developed in 1977, was called “Northern blotting” by James Alwine, David Kemp and George Stark at Stanford University as a play on Southern blotting (3). During a quick chat, Nowinski and Burnette decided to continue the directional joke. They dubbed Burnette’s method “Western blotting” simply because the laboratory was located on the West Coast.

As Burnette was developing his technique, a paper appeared in Proceedings of the National Academy of Sciences that described a similar approach (4). But Burnette was convinced that his method made it easier to transfer the proteins from gel to membrane, get antibody detection and analyze the blot. So he began to put together a manuscript. At this point, Nowinski says he told Burnette, “I didn’t think it would be appropriate for me to come on the paper as an author, because it was really all his work.”

When Burnette, the sole author of the manuscript, sent his work to Analytical Biochemistry, it was promptly rejected. The rejection wasn’t because of the method’s similarity to the technique in the PNAS paper but because it seemed pedestrian, and one reviewer had taken particular offense to the whimsical name.

On receiving the rejection, “I thought, ‘What the heck,’ and didn’t pay much attention,” says Burnette. But he had given preprints of the paper to his friends. They photocopied the paper and gave it to their friends, who repeated the process. “Pretty soon, I was running a daily seminar on blotting by telephone. I was talking to everyone on the planet who was trying to reach me because they couldn’t read the Xeroxed copy they had,” says Burnette. “I had moved to the Salk Institute from Fred Hutchinson by then. [Western blotting] was taking up all my time by talking on the phone.”

Frustrated, Burnette called back the editors of Analytical Biochemistry. “I told them, ‘This is crazy. Everybody knows about this technique now that I’ve not published for two years. You think you might like to publish it now?’” The journal at this point agreed and published the paper in 1981. “Then I got deluged with reprint requests!” says Burnette.

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COMMENTS:

Nice article. So, is it "Western" or "western"? Or does either work, without a preference? - Bruce Maryanoff (bmaryano@scripps.edu)

 

I was a high school friend of Neil's and recall recognizing how committed he was to acting as we enjoyed a coke one evening at local hangout and he still had part of his make up on from a dress rehersal he had just attended. Little did I know how focused he would become on a wide range of endeavors or how greatly he would apply himself to so very many. I next met up with him at a 40th reunion and was thoroughly impressed by the reach of his shift to scientific discipline. At the time he was a full colonel filling a general's slot in the US Army. Congratulations to Neil on a life well lived and leaving a trail of useful accomplishments. And thanks to the author for seeing that such a life does not escape notice. James Sizemore - Bedford, TX.

 

I went to high school with Neil and we were good friends. After going our separate ways I lost touch with him. Now that we are retired I located him via facebook, and retrieved this article via that link. I used the western blot method while I was teaching and never knew it had a connection to an old buddy. I find the article fascinating, well written and a worthy tribute to someone who has so effectively contributed to scientific knowledge. I wish to thank the author for giving credit where credit was obviously due. A Cheer for Neil! With best regards, Gil R Alexander, Retired Director of the Montana Science Institute, and former President of the Montana Academy of Sciences

 

1 Comments

  • Neal: I just received an e-mail from Jim Kaper in which he said he and his wife had just returned from India with a visit to the Taj Mahal where he and I bumped into each other in 1990 by accident (we were there for different meetings). I was thus responding to him that you and I had visited there in 1992 and had our picture taken in front of the same minaret, except my picture with him was from 15 feet away since the place was crowded with 1000s of people whereas our picture was taken from over 100 feet away since we visited during a curfew where we had the whole place to ourselves with nobody but us and our guides! The two pictures are still on my bulletin board in my office to remind me of happy days and good friends. Hope all is well. Best regards, Roy

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