There was "standing room only" at the beginning of this far ranging session that was sponsored jointly by the ABRF and the ASBMB Educational Affairs Committee. As summarized in the opening statement by Ken Williams, who chaired this session, recent improvements in protein and nucleic acid sequencing, synthesis, and analysis techniques and instrumentation have brought about a "virtual renaissance in biotechnology rivaling that which has occurred in molecular biology. The degree of excitement in biotechnology has been pushed to even new heights because of the parallel establishment of more than 200 core facilities who are dedicated to bringing these new technologies to bear on important biological problems. The fact that nearly every investigator now has access to amino acid analysis, protein sequencing, peptide synthesis, and oligonucleotide synthesis undoubtedly helped to account for the high level of interest in this symposium.
The first and last speakers, Dave Speicher and Ron Niece, respectively, provided a realistic assessment of the capabilities of "state-of-the-art" biotechnology that was firmly based on the analysis of data from past ABRF studies involving standard amino acid, peptide, and protein samples as well as on surveys completed by 124 ABRF member laboratories. As promised, the remaining six speakers provided a glimpse of the future as it already exists in their own laboratories. Kathy Stone (Yale U.) provided details on an in situ cyanogen bromide digestion/extraction procedure that, based on studies on 11 standard proteins, can be used to elute an average of 74% of PVDF-blotted, Coomassie Blue-stained proteins in a form that is suitable for subsequent trypsin digestion and narrow bore HPLC separation. Donald Hunt (University of Virginia)
documented recent advances in mass spectrometry which permit his laboratory to sequence most of the peptides obtained from tryptic digests that are carried out on as little as 5-10 pmol of protein. Ion trap detectors, capable of detecting as few as 25 ions, should soon push these sensitivity limits even lower. James Bowie's (UCLA) novel approach of asking, "What sequences can fold into a given structure?", as opposed to the more traditional, "What structure does a given sequence fold into?", provided the starting point for the development of more powerful approaches for uncovering functional domains in proteins. Automated DNA sequencing has advanced to the point that Craig Venter's group (NIH), is now able to read as many as 700 bases from a single run and, with the aid of three technicians, his laboratory can now sequence as much as 48 kb/day. Recent improvements in robotics promise to significantly decrease the amount of "human intervention" that is now required in automated DNA sequencing. Richard Houghten (Torrey Pines Institute for Molecular Studies) provided an overview of multiple peptide synthesis strategies which hold the promise of meeting the ever increasing demand for synthetic peptides. Finally, Michael Ng (Stanford U.) described techniques based on continuous zone electrophoresis (CZE) which permit as little as 1000 molecules of an individual component, which may be isolated from a single cell, to be analyzed.
Since less than 20% of ABRF Core Facilities are currently equipped for automated DNA sequencing, mass spectrometry, or CZE, one of the key messages from this symposium was that increased funding and support of core facilities will surely be required in order to bring these "theoretical" advances within everyday reach of most investigators.
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Created: 21st August 1995
Last modified: 21st August 1995