We've crystallized a complex of an Fab bound to a protein. We have the hybridomas from which the Fab was prepared, but no protein sequence for the antibody. We're trying to plot the easiest course to get the sequence (since the crystals, alas, do not diffract to sufficiently high resolution so as to allow us to read off the sequence from the map).
Since we have access to a lot of pure protein, I wonder if some clever mass spec jock would be able to assemble enough overlapping sequenced fragments so as to give complete coverage of the protein. Has anyone done this/had this done for them? Alternatively, I guess we can reverse transcribe message from the hybridoma cells, but I've heard suggestions that this is not necessarily straightforward (e.g., hybridomas may contain "rogue" Ig mRNA that will show up in a PCR experiment), so I'm hoping to avoid this particular can of worms. Any insights welcome. Cheers, Pat --------------------------------------------------------------------------------------- Patrick J. Loll, Ph. D. Professor of Biochemistry & Molecular Biology Director, Biochemistry Graduate Program Drexel University College of Medicine Room 10-102 New College Building 245 N. 15th St., Mailstop 497 Philadelphia, PA 19102-1192 USA (215) 762-7706 pat.l...@drexelmed.edu
