The Synergism of Protein Chemistry and Recombinant DNA Techniques

This paper will attempt to provide an overview as to how recombinant DNA and protein chemical techniques work hand-in-glove in order to relatively easily determine complete structural information concerning a series of molecules. Emphasis will be placed on immunoglobulin sequencing utilizing picomole amounts of murine and human hybridomas to determine the N-terminal one third of both heavy and light polypeptide chains and the use of oligonucleotide primers for mRNA sequencing of the same molecules. The overlaps in the center of the variable regions provide assurance that the two techniques allow the complete primary structure of immunoglobulin variable regions to be deduced rather rapidly. Techniques for fragmentation are unnecessary (which would be required if only protein chemistry techniques were used) and second and third “primings” are not required (as would be necessitated by relying solely on mRNA sequencing techniques). These studies have resulted in the complete structural analysis of a series of human and murine hybridomas from extremely small amounts of ascites fluid, culture fluid and cell pellets.