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Biotech / Medical : HuMAB companies -- Ignore unavailable to you. Want to Upgrade?

To: Icebrg who wrote (649)	6/14/2004 5:00:05 PM
From: tuck	Respond to of 1022

[Anchored periplasmic expression, a versatile technology for the isolation of high-affinity antibodies from Escherichia coli-expressed libraries] >>Published online before print June 14, 2004 Proc. Natl. Acad. Sci. USA, 10.1073/pnas.0400187101 Applied Biological Sciences Anchored periplasmic expression, a versatile technology for the isolation of high-affinity antibodies from Escherichia coli-expressed libraries Barrett R. Harvey , George Georgiou , Andrew Hayhurst , Ki Jun Jeong , Brent L. Iverson ¶, and Geoffrey K. Rogers Institute for Cellular and Molecular Biology and Departments of Chemical Engineering, ¶Chemistry and Biochemistry, and Biomedical Engineering, University of Texas, Austin, TX 78712 Edited by James A. Wells, Sunesis Pharmaceuticals, Inc., South San Francisco, CA, and approved May 10, 2004 (received for review January 9, 2004) Anchored periplasmic expression (APEx) is a technology for the isolation of ligand-binding proteins from combinatorial libraries anchored on the periplasmic face of the inner membrane of Escherichia coli. After disruption of the outer membrane by Tris-EDTA-lysozyme, the inner-membrane-anchored proteins readily bind fluorescently labeled ligands as large as 240 kDa. Fluorescently labeled cells are isolated by flow cytometry, and the DNA of isolated clones is rescued by PCR. By using two rounds of APEx, the affinity of a neutralizing antibody to the Bacillus anthracis protective antigen was improved >200-fold, exhibiting a final KD of 21 pM. This approach has several technical advantages compared with previous library screening technologies, including the unique ability to screen for ligand-binding proteins that bind endogenously expressed ligands fused to a short-lived GFP. Further, APEx is able to display proteins either as an N-terminal fusion to a six-residue sequence derived from the native E. coli lipoprotein NlpA, or as a C-terminal fusion to the phage gene three minor coat protein of M13. The latter fusions allow hybrid phage display/APEx strategies without the need for further subcloning.<< Cheers, Tuck

To: Icebrg who wrote (649)	6/22/2004 11:30:06 AM
From: Icebrg	Read Replies (1) \| Respond to of 1022

Curagen and Seattle Genetics. Interesting to note from today's PR that the cooperation between CRGN and ABGX seems to have resulted in something workable. I guess IMGN feels a little bit pi**sed off today as they are supposed to be ABGX's partner for this type of mAb-enhancement. Although we have not yet seen any results yet despite Abgenix having a head-start of abt. three years. As a matter of fact Withy recently said, that they were not going to pursue any development of mAb-conjugates unless they are able to partner the program. Erik