From owner-molreps@hgmp.mrc.ac.uk Mon Oct 16 14:33:11 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 3880317B4D; Mon, 16 Oct 2000 14:33:10 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 60001) id 9CDDC17B4C; Mon, 16 Oct 2000 14:33:06 +0100 (BST) To: molreps@net.bio.net Newsgroups: bionet.molecules.repertoires From: pcxsc@nottingham.ac.uk (Simon Cross) Subject: Phage Display Date: 27 Sep 2000 14:27:32 +0100 Organization: ACS, The University of Nottingham Message-ID: <39D1F4C1.88CC4651@nottingham.ac.uk> Sender: owner-molreps@hgmp.mrc.ac.uk Precedence: bulk 1) Does anyone know the best newsgroup for phage display? 2) I am using the Griffin1 library of phage displayed scFv's and can never seem to get past the soluble fragment stage. I get very few positives after 4 rounds of screening the phage, and I get reproducible results of the clonal phage specifically binding my hapten molecule shown by ELISA. However, when I express the soluble antibody from these positives I never get the scFv binding in the same way (although it's definitely expressed ok, shown by dot blot). I have heard of problems with naive libraries and am aware that the scFv may not fold in the same way when fused to the g3 coat protein, but there are papers out there saying that this technique works! I would appreciate any suggestions, Simon pcxsc@nottingham.ac.uk --- From owner-molreps@hgmp.mrc.ac.uk Mon Oct 16 14:34:08 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id CF80B17B50; Mon, 16 Oct 2000 14:34:07 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 60001) id 2476917B52; Mon, 16 Oct 2000 14:34:05 +0100 (BST) To: molreps@net.bio.net Newsgroups: bionet.molecules.repertoires From: mrc7@cam.ac.uk (Mike Clark) Subject: Re: Phage Display Date: 27 Sep 2000 14:15:33 +0100 Organization: Cambridge University, Department of Pathology. Message-ID: References: <39D1E671.809A818B@nottingham.ac.uk> Sender: owner-molreps@hgmp.mrc.ac.uk Precedence: bulk In article <39D1E671.809A818B@nottingham.ac.uk>, Simon Cross wrote: > 1) Does anyone know the best newsgroup for phage display? > > 2) I am using the Griffin1 library of phage displayed scFv's and can > never seem to get past the soluble fragment stage. I get very few > positives after 4 rounds of screening the phage, and I get reproducible > results of the clonal phage specifically binding my hapten molecule > shown by ELISA. However, when I express the soluble antibody from these > positives I never get the scFv binding in the same way (although it's > definitely expressed ok, shown by dot blot). I have heard of problems > with naive libraries and am aware that the scFv may not fold in the same > way when fused to the g3 coat protein, but there are papers out there > saying that this technique works! I would appreciate any suggestions, > > Simon > pcxsc@nottingham.ac.uk > > Simon, I can't answer your specific questions but I suggest that you try the bionet.molecules.repertoires newsgroup (I have cross posted this reply). One of the problems with published scientific techniques is that whilst reports of success are readily published, reports of failure are not! Mike Clark, -- o/ \\ // || ,_ o M.R. Clark, PhD. Division of Immunology <\__,\\ // __o || / /\, Cambridge University, Dept. Pathology "> || _`\<,_ // \\ \> | Tennis Court Rd., Cambridge CB2 1QP ` || (_)/ (_) // \\ \_ Tel.+44 1223 333705 Fax.+44 1223 333875 --- From owner-molreps@hgmp.mrc.ac.uk Fri Oct 27 09:48:01 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id DA04717B26; Fri, 27 Oct 2000 09:48:00 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 60001) id BAE2117B2A; Fri, 27 Oct 2000 09:47:58 +0100 (BST) To: molreps@net.bio.net Newsgroups: bionet.molecules.repertoires From: soldatov@molgen.mpg.de (Aleksey Soldatov) Subject: Buffers and programs on http://www.molbiol.edu.ru/eng Date: 26 Oct 2000 16:54:02 +0100 Organization: MPI Message-ID: <39F84C5A.F43EFD4E@molgen.mpg.de> Sender: owner-molreps@hgmp.mrc.ac.uk Precedence: bulk A lot of useful resources on http://www.molbiol.edu.ru/eng In "Protocols" section you will find a large list of recipes of salt solutions, laboratory buffers, media, antibiotics etc. http://www.molbiol.edu.ru/eng/protocol In "Calculations" section there are a number of small programs, which will help in common laboratory calculations (weight - molar conversion for proteins and nucleic acids; [krpm] - [kg] conversion for centrifugation; estimation of time for bacteria growth; spectrophotometric measurement of nucleic acids concentration and so on). http://www.molbiol.edu.ru/eng/scripts BioMail is a program that simplifies and automates searching for biological and medical literature. BioMail will search PubMed, the largest publically available online database for biomedical journals, using your search terms. BioMail then e-mails you the results weekly, biweekly or monthly as you choose. http://molbiol.edu.ru/cgi-bin/biomail/users.pl The site is predominantly in Russian. Here we translated only the pages, for which we did not find English analogues. Aleksey ---