From owner-repertoires@net.bio.net Sun Oct 06 23:00:00 1996 Path: biosci!agate!news.ucdavis.edu!info.ucla.edu!unixg.ubc.ca!lpss From: lpss@unixg.ubc.ca (Alex Chang) Newsgroups: bionet.molecules.repertoires Subject: What is the best immuno-panning method Date: 7 Oct 1996 18:28:26 GMT Organization: University of British Columbia, Vancouver, B.C., Canada Lines: 11 Message-ID: <53bi4a$5gh@nntp.ucs.ubc.ca> NNTP-Posting-Host: netinfo.ubc.ca X-Newsreader: TIN [version 1.2 PL2] I recently have lots of trouble to do immuno-panning using MaxiSorb Immunotube (Nunce) for my phage (M13) display library. I'd like to know whether there is a better method to do this. -- Alex Chang Pathology University of British Columbia achang@hivnet.ubc.ca From owner-repertoires@net.bio.net Tue Oct 08 23:00:00 1996 Path: biosci!daresbury!not-for-mail From: "Janet Clench, Library, Tel:(39 6)91093220" Newsgroups: bionet.molecules.repertoires Subject: the latest update on COmb. Libraries and similar repertoires Date: 9 Oct 1996 08:36:49 +0100 Lines: 221 Sender: lpddist@mserv1.dl.ac.uk Distribution: bionet Message-ID: <53fkmh$9la@mserv1.dl.ac.uk> Original-To: molreps@dl.ac.uk ******************************************************** SUBJECT: Combinatorial & Phage Libraries DATE: September 23, 30, October 7 1996 ******************************************************** Biochemistry 35: 36 (SEP 10 1996) MA Kelly, HB Liang, II Sytwu, I Vlattas, NL Lyons, BR Bowen, LP Wennogle Characterization of SH2 - Ligand interactions via library affinity selection with mass spectrometric detection 11747-11755 Journal of Biological Chemistry 271: 37 (SEP 13 1996) MJ Rosok, DE Yelton, LJ Harris, J Bajorath, KE Hellstrom, I Hellstrom, GA Cruz, K Kristensson, H Lin, WD Huse, SM Glaser A combinatorial library strategy for the rapid humanization of anticarcinoma BR96 Fab 22611-22618 Protein Science 5: 9 (SEP 1996) ZJ Ren, GK Lewis, PT Wingfield, EG Locke, AC Steven, LW Black Phage display of intact domains at high copy number: A system based on SOC, the small outer capsid protein of bacteriophage T4 1833-1843 Chemistry & Biology 3: 8 (AUG 1996) SB Feng, TM Kapoor, F Shirai, AP Combs, SL Schreiber Molecular basis for the binding of SH3 ligands with non-peptide elements identified by combinatorial synthesis 661-670 Chemistry & Biology 3: 8 (AUG 1996) HM Geysen, CD Wagner, WM Bodnar, CJ Markworth, GJ Parke, FJ Schoenen, DS Wagner, DS Kinder Isotope or mass encoding of combinatorial libraries 679-688 Journal of Organic Chemistry 61: 18 (SEP 6 1996) YF Shao, WC Still Sequence-selective receptors of peptides. A simple molecular design for construction of large combinatorial libraries of receptors 6086-6087 Journal of Immunology 157: 3 (AUG 1 1996) HI Yamanaka, T Inoue, O Ikedatanaka Chicken monoclonal antibody isolated by a phage display system 1156-1162 Current Opinion in Structural Biology 6: 4 (AUG 1996) ZX Shao, FH Arnold Engineering new functions and altering existing functions 513-518 Nature Biotechnology 14: 9 (SEP 1996) S Klussmann, A Nolte, R Bald, VA Erdmann, JP Furste Mirror-image RNA that binds D-adenosine 1112-1115 Nature Biotechnology 14: 9 (SEP 1996) BT Mcguinness, G Walter, K Fitzgerald, P Schuler, W Mahoney, AR Duncan, HR Hoogenboom Phage diabody repertoires for selection of large numbers of bispecific antibody fragments 1149-1154 Journal of Molecular Biology 262: 1 (SEP 13 1996) YG Mikawa, IN Maruyama, S Brenner Surface display of proteins on bacteriophage lambda heads 21-30 Journal of Biomolecular NMR 8: 1 (JUL 1996) G Barbato, DO Cicero, E Bianchi, A Pessi, R Bazzo High-resolution solution structure of two members of a conformationally homogeneous combinatorial peptide library based on the classical zinc-finger motif 36-48 Nature Medicine 2: 9 (SEP 1996) RHJ Begent, MJ Verhaar, KA Chester, JL Casey, AJ Green, MP Napier, LD Hopestone, N Cushen, PA Keep, CJ Johnson, RE Hawkins, AJW Hilson, L Robson Clinical evidence of efficient tumor targeting based on single-chain Fv antibody selected from a combinatorial library 979-984 Tetrahedron 52: 37 (SEP 9 1996) IE Pop, CF Dhalluin, BP Deprez, PC Melnyk, GM Lippens, AL Tartar Monitoring of a three-step solid phase synthesis involving a Heck reaction using magic angle spinning NMR spectroscopy 12209-12222 Tetrahedron Letters 37: 36 (SEP 2 1996) SR Gilbertson, XF Wang The combinatorial synthesis of chiral phosphine ligands 6475-6478 Tetrahedron Letters 37: 36 (SEP 2 1996) KW Jung, XY Zhao, KD Janda A linker that allows efficient formation of aliphatic C-H bonds on polymeric supports 6491-6494 Brazilian Journal of Medical and Biological Research 29: 9 (SEP 1996) R Pasqualini, E Koivunen, E Ruoslahti Peptides in cell adhesion: Powerful tools for the study of integrin-ligand interactions 1151-1158 Journal of Clinical Investigation 98: 4 (AUG 15 1996) HK Jin, B Li, B Cunningham, J Tom, RH Yang, P Sehl, GR Thomas, A Ko, D Oare, DG Lowe Novel analog of atrial natriuretic peptide selective for receptor-A produces increased diuresis and natriuresis in rats 969-976 Immunology 89: 1 (SEP 1996) M Duenas, LT Chin, AC Malmborg, R Casalvilla, M Ohlin, CAK Borrebaeck In vitro immunization of naive human B cells yields high affinity immunoglobulin G antibodies as illustrated by phage display 1-7 Trends in Biotechnology 14: 9 (SEP 1996) MA Jongsma, WJ Stiekema, D Bosch Combatting inhibitor-insensitive proteases of insect pests 331-333 Trends in Biotechnology 14: 9 (SEP 1996) C Khosla, RJX Zawada Generation of polyketide libraries via combinatorial biosynthesis 335-341 Archives of Biochemistry and Biophysics 333: 1 (SEP 1 1996) SL Deutscher, ME Crider, JA Ringbauer, AA Komissarov, TP Quinn Stability studies of nucleic acid-binding Fab isolated from combinatorial bacteriophage display libraries 207-213 Biochimica et Biophysica Acta - Molecular Basis of Disease 1316: 3 (AUG 23 1996) S Tyutyulkova, QS Gao, A Thompson, S Rennard, S Paul Efficient vasoactive intestinal polypeptide hydrolyzing autoantibody light chains selected by phage display 217-223 European Journal of Biochemistry 240: 1 (AUG 15 1996) B Fleckenstein, H Kalbacher, CP Muller, D Stoll, T Halder, G Jung, KH Wiesmuller New ligands binding to the human leukocyte antigen class II molecule DRB1(*)0101 based on the activity pattern of an undecapeptide library 71-77 Angewandte Chemie - International Edition in English 35: 15 (AUG 19 1996) BM Cole, KD Shimizu, CA Krueger, JPA Harrity, ML Snapper, AH Hoveyda Discovery of chiral catalysts through ligand diversity: Ti-catalyzed enantioselective addition of TMSCN to meso epoxides 1668-1671 International Journal of Peptide and Protein Research 48: 2 (AUG 1996) AM Sefler, G Lauri, PA Barlett A convenient method for determining cyclic peptide conformation from 1D H-1-NMR information 129-138 Journal of the American Chemical Society 118: 33 (AUG 21 1996) YH Chu, YM Dunayevskiy, DP Kirby, P Vouros, BL Karger Affinity capillary electrophoresis mass spectrometry for screening combinatorial libraries 7827-7835 Journal of the American Chemical Society 118: 34 (AUG 28 1996) BA Katz, BS Liu, R Cass Structure-based design tools: Structural and thermodynamic comparison with biotin of a small molecule that binds to streptavidin with micromolar affinity 7914-7920 Journal of the American Chemical Society 118: 34 (AUG 28 1996) K Russell, DC Cole, FM Mclaren, DE Pivonka Analytical techniques for combinatorial chemistry: Quantitative infrared spectroscopic measurements of deuterium-labeled protecting groups 7941-7945 Journal of Organic Chemistry 61: 16 (AUG 9 1996) R Fathi, MJ Rudolph, RG Gentles, R Patel, EW Macmillan, MS Reitman, D Pelham, AF Cook Synthesis and properties of combinatorial libraries of phosphoramidates 5600-5609 Tetrahedron Letters 37: 35 (AUG 26 1996) P Seneci, C Sizemore, K Islam, P Kocis Combinatorial chemistry and natural products. Teicoplanin aglycone as a molecular scaffold for solid phase synthesis of combinatorial libraries 6319-6322 Drug Discovery Today 1: 9 (SEP 1996) N Terrett Combinatorial chemistry 402 Research on Chemical Intermediates 22: 7 (1996) I Ugi, M Goebel, B Gruber, M Heilingbrunner, C Heiss, W Horl, O Kern, M Starnecker, A Domling Molecular libraries in liquid phase via UGI-MCR 625-644 Theoretica Chimica Acta 94: 2 (AUG 1996) S Fujita Pseudo-point groups and chronality in enumeration of reaction pairs 105-124 Chemicke Listy 90: 8 (AUG 1996) F Svec Combinatorial syntheses or ''mathematics'' and ''libraries'' in chemistry 477-485 From owner-repertoires@net.bio.net Wed Oct 16 23:00:00 1996 Path: biosci!fcs280s.ncifcrf.gov!not-for-mail From: toms@ncifcrf.gov (Tom Schneider) Newsgroups: bionet.info-theory,bionet.molecules.repertoires Subject: Re: General Questions (Observations to follow) Date: 17 Oct 1996 17:12:04 GMT Organization: NCI-FCRDC Frederick Biomedical Supercomputing Center Lines: 51 Message-ID: <545pd4$9a24@ncisun1-nf0.ncifcrf.gov> References: <5413ai$9a23@ncisun1-nf0.ncifcrf.gov> NNTP-Posting-Host: kaylor.ncifcrf.gov X-Newsreader: TIN [UNIX 1.3 950824BETA PL0] Xref: biosci bionet.info-theory:4333 bionet.molecules.repertoires:257 Ben Lieberman (ben.lieberman@uchsc.edu) wrote: : Already figured out a way to do it in-vivo. It would require modifying the : host strain to protect internal sequences against digestion. This may be : possible by "evolving" a methylase first, then going after the restriction : enzyme, or by "co-evolving" the host and the enzyme (just like nature only : faster). The methylase method would reduce the number of possible : sequences to be selected to those containing methylation (A or C) residues. It's not clear to me how this selection would work. (You might not want to post your idea; it may be patentable.) Your discussion above suggests a scheme. Possibly one could put the initial methylase/restriction enzyme on a plasmid to allow rounds of dirty PCR amplification to make mutations. (The plasmid replication and drug resistance would be easily selected for, bad ones would not replicate.) Then select bacteria that are resistant to a lambda vir (ie cI- operator-) that has many copies of the sequence you want to attack. One might think that the main problem would be detecting bacteria that become resistant to lambda infection, but if you did rounds of extraction of the plasmid, PCR mutation and re-transformation, this would be avoided! The sequence you have in the lambda should initially be similar to the methylase/restriction system. : Alternatively there may be a possible way of combinatorial combinations of : point directed mutagensis to do the same in vitro - but I have to think : more about it. The possibility exists to express a restriction enzyme on the surface of a bacteriophage, along with a streptavidin to bind the phage to the end of a repeat-sequence DNA attached to a magnetic bead. Phage that consistently cut themselves away from the bead would be the ones you want. I'm cross posting to bionet.molecules.repertoires. Lest folks think that this is not relevant to the bionet.info-theory group, consider that the evolution of restriction enzyme binding sites is MORE mysterious than that of "fuzzy" sites like ribosome or spliceosome sites because of Shannon's channel capacity precision theorem. How can they be so precise without having many "pins"? How can they evolve across the big gaps in sequence patterns? : Thanks for the reply. You're welcome! Tom Schneider National Cancer Institute Laboratory of Mathematical Biology Frederick, Maryland 21702-1201 toms@ncifcrf.gov http://www-lmmb.ncifcrf.gov/~toms/ From owner-repertoires@net.bio.net Mon Oct 21 23:00:00 1996 Path: biosci!daresbury!not-for-mail From: Andrew Wallace Newsgroups: bionet.molecules.repertoires Subject: Re: General Questions (Observations to follow) Date: 22 Oct 1996 18:57:24 +0100 Lines: 57 Sender: lpddist@mserv1.dl.ac.uk Distribution: bionet Message-ID: <54j1u4$jom@mserv1.dl.ac.uk> Original-Sender: awallace@uk.ac.qub MIME-Version: 1.0 X-Authentication: none Read-Receipt-To: Andrew Wallace Original-To: molreps@dl.ac.uk Reposting this as it was sent to the wrong address the first time... On Fri, 18 Oct 1996 12:55:04 +0100 Andrew Wallace wrote: > > Dear Tom et al., > > Would it be possible for one of you to cross-post the start of this > discussion to bionet.molecules.repertoires so that everyone can follow > it from the beginning? > > Thanks, > > Andrew > > P.S. Note new web page location (see sig) and new direct phone number > Tel. +44-1232-272089/2196 (office/lab) > > On 17 Oct 1996 17:12:04 GMT Tom Schneider wrote: > > > > Ben Lieberman (ben.lieberman@uchsc.edu) wrote: > > > > : Already figured out a way to do it in-vivo. It would require modifying the > > : host strain to protect internal sequences against digestion. This may be > > : possible by "evolving" a methylase first, then going after the restriction > > : enzyme, or by "co-evolving" the host and the enzyme (just like nature only > > : faster). The methylase method would reduce the number of possible > > : sequences to be selected to those containing methylation (A or C) residues. > > > > It's not clear to me how this selection would work. (You might not want to > > post your idea; it may be patentable.) > > > > [I deleted the rest to save bandwidth...] > > > Tom Schneider > > National Cancer Institute > > Laboratory of Mathematical Biology > > Frederick, Maryland 21702-1201 > > toms@ncifcrf.gov > > http://www-lmmb.ncifcrf.gov/~toms/ > > > > > > ------------------------------------------------------------------ > Andrew Wallace, Ph.D, Queen's University Belfast, N. Ireland (UK) > a.wallace@qub.ac.uk http://web.qub.ac.uk/bb/awpage/wallace.html > ------------------------------------------------------------------ Andrew Wallace, Ph.D, Queen's University Belfast, N. Ireland (UK) a.wallace@qub.ac.uk http://web.qub.ac.uk/bb/awpage/wallace.html From owner-repertoires@net.bio.net Tue Oct 22 23:00:00 1996 Path: biosci!EMAIL.UNC.EDU!briankay From: briankay@EMAIL.UNC.EDU (Brian Kay) Newsgroups: bionet.molecules.repertoires Subject: Phage Display Book out Date: 23 Oct 1996 11:45:47 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 24 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Hi All, The "Phage Display of Peptides and Proteins: A Laboratory Manual" has finally been published. You can find the table of contents at URL: http://www.unc.edu/depts/biology/bkay/phagedisplay.html Brian =-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-==-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-= Brian Kay Associate Professor of Biology 321 Fordham Hall University of North Carolina at Chapel Hill Chapel Hill, NC 27599-3280 Office: 919-962-2144 Fax: 919-962-1625 Email: briankay@email.unc.edu Webpages: http://kay02.bio.unc.edu/ Phage Display Book: http://www.unc.edu/depts/biology/bkay/phagedisplay.html =-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-==-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-= From owner-repertoires@net.bio.net Tue Oct 22 23:00:00 1996 Path: biosci!daresbury!bioftp.unibas.ch!infobiogen.fr!jussieu.fr!univ-lyon1.fr!howland.erols.net!EU.net!newsfeed.internetmci.com!csn!nntp-xfer-1.csn.net!news-2.csn.net!csn!nntp-xfer-2.csn.net!tali.UCHSC.edu!ben.lieberman From: ben.lieberman@uchsc.edu (Ben Lieberman) Newsgroups: bionet.molecules.repertoires Subject: Re: General Questions (Observations to follow) Date: Wed, 23 Oct 1996 08:49:04 -0600 Organization: UCHSC Lines: 39 Distribution: bionet Message-ID: References: <54j1u4$jom@mserv1.dl.ac.uk> NNTP-Posting-Host: brbmxt204.uchsc.edu Mime-Version: 1.0 Content-Type: text/plain; charset=ISO-8859-1 Content-Transfer-Encoding: 8bit X-Newsreader: Yet Another NewsWatcher 2.3.1 In article <54j1u4$jom@mserv1.dl.ac.uk>, Andrew Wallace wrote: > > Would it be possible for one of you to cross-post the start of this > > discussion to bionet.molecules.repertoires so that everyone can follow > > it from the beginning? > > > > Thanks, > > > > Andrew Sure thing Andrew, This disscussion started with a list of questions I had about information theory and biology. You can probably still find the beginning of the thread in the bionet.info-theory group. The general gist of this part of the thread was about how to "co-evolve" bactera (e.g. E. coli) to produce a restriction enzyme that would specifically recognize any sequence you were interested in cutting. My rough idea was to start with a mutator strain of E.coli (one in which the DNA polymerase error correction activity is comprimised) and a plasmid encoding a "randomized" version of a well known restriction enzyme (e.g. EcoRI) were the DNA contacts are well established. By allowing the amino acids comprising the recognition site in the protein to be altered (yes the mutator strain will also make hash of the coding sequence, but this is going to be a selection screen so it won't matter), and providing a the sequence of interest in a selectable way (Tom suggested using lambda phage), you can allow a natural selection to take place. The details of this are very shaky, and I am not sure yet how to pull it off, but the value (commercial and scientific) are obvious. Think about how much restriction enzyme is produced by companies on a yearly basis and you get the idea. Also, the idea of engineering DNA binding proteins to suit is very attractive. Ben Lieberman ben.lieberman@uchsc.edu -- I sometimes wish that things could be more like they are. - J. Jensen - From owner-repertoires@net.bio.net Sun Oct 27 22:00:00 1996 Path: biosci!internet!biosci!not-for-mail From: biohelp (BIOSCI Administrator) Newsgroups: bionet.molecules.repertoires Subject: BIOSCI/bionet miniFAQ & Fundraiser Date: 28 Oct 1996 02:00:36 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 239 Sender: daemon@net.bio.net Distribution: world Message-ID: <199610281000.CAA10608@net.bio.net> NNTP-Posting-Host: net.bio.net (LAST REVISION: 30-JUL-95) This BIOSCI "miniFAQ" is designed to answer the questions that come up the *most frequently*. The main BIOSCI FAQ (Frequently Asked Questions) is accessible on the World Wide Web at URL http://www.bio.net/. If you can not find an answer to your question in this or other documentation, the BIOSCI technical support staff answers e-mail queries sent to biosci-help@net.bio.net We can only answer questions about the use of the newsgroups and mailing lists. We unfortunately do not have the staff to do Internet information searches or answer scientific questions. Please post those to the appropriate BIOSCI/bionet newsgroups. Contents: -------- 0) BIOSCI NEEDS YOUR SUPPORT!! 1) Using the WWW to access the BIOSCI/bionet newsgroups. 2) What to do about "spams," i.e., junk mail, ads, etc. 3) Examples of subscribing and unsubscribing to the mailing lists. 4) The BIOSCI user address and research interest directory. 0) BIOSCI NEEDS YOUR SUPPORT!! ------------------------------ BIOSCI's government funding has been expended, and we are now operating solely from advertising revenue that we have raised from our Web site at http://www.bio.net/. We need just a few minutes of your time to help us serve you. You can do two important things which will take very little time for you individually and will immensely help us continue to help you. First, please use our WWW system at http://www.bio.net/ to access the archives. You can post or reply to messages via your Web browser as described in item #1 below. Your usage helps attract sponsors. If you contact any of our sponsors, please be sure to thank them for supporting BIOSCI. It is critical for them to get this feedback if they are to continue their sponsorship for the long term. Second, if you work for a company or organization that provides products or services of interest to the biology community, please pass this message on to your marketing or marketing communications department or other appropriate group. Please ask them to help support BIOSCI by sponsoring our Web site and explain the uses and benefits of the system to the biology community. If they are interested, they can then contact us for further information at our tech support address, biosci-help@net.bio.net. 1) Using the WWW to access the BIOSCI/bionet newsgroups. -------------------------------------------------------- As of 10 December 1995, all BIOSCI/bionet full newsgroups are accessible through the World Wide Web (WWW) at URL http://www.bio.net. One can read and reply publicly or privately to both recent postings and archived messages through one's Web browser if it is configured properly to send e-mail. Each newsgroup is equipped with its own WAIS index. 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Sincerely, Dave Kristofferson BIOSCI/bionet Manager biosci-help@net.bio.net From owner-repertoires@net.bio.net Sun Oct 27 22:00:00 1996 Path: biosci!rutgers!news.sgi.com!news.sprintlink.net!news-peer.sprintlink.net!arclight.uoregon.edu!news.uoregon.edu!news.rediris.es!news.csic.es!news From: cibsm1s@fresno.csic.es (Jesus Sanz) Newsgroups: bionet.molecules.repertoires Subject: Peptide display Date: 28 Oct 1996 17:34:35 GMT Organization: CIB/CSIC Lines: 37 Message-ID: <552qrb$q3p@granado> NNTP-Posting-Host: jesus.cib.csic.es X-Newsreader: WinVN 0.92.6+ Hi, netters, I've recently used the phage display system and was able to identify a family of sequence-related peptides that bind to my protein. I syntethized the peptides but they did not have any effect on the activity of the enzyme. However, I constructed a fusion with a choline-binding protein that can be purified in a single step in DEAE-cellulose. The reason for doing this was that maybe the peptide needed to be conformationally constrained in order to interact with the enzyme (in the phage-display system the peptide is actually fused to gene III protein). Such fusion works, i.e., modulates the activity of the enzyme, although the activation is not what I call "impressing". I am sure that, if I can find the suitable "scaffold", the peptide fused to it can be extremely effective. I would appreciate any information from you about polypeptides with some of all of the following properties: 1) Allow fusions 2) Overproduced and easy to purify 3) Small size (<12 kDa, so that I can monitor the binding by NMR techniques, for example) 4) Non toxic for humans (so that I can use them "in vivo") Am I asking for too much? (Don't answer me). Thank you very much for any response. Jesus --------- Dr. Jesus M. Sanz E-mail:cibsm1s@fresno.csic.es Centro de Investigaciones Talk: jesuso@jesus.cib.csic.es Biologicas Phone:(34)(1)5611800 Ext 4368 Velazquez, 144; 28006-Madrid, Spain Fax:(34)(1)5627518 --------- From owner-repertoires@net.bio.net Tue Oct 29 22:00:00 1996 Path: biosci!NMSU.EDU!dkim From: dkim@NMSU.EDU ("D. KIM") Newsgroups: bionet.molecules.repertoires Subject: ultra pellet of M13 Date: 30 Oct 1996 06:27:01 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 5 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Can anyone tell me what are good conditions for doing an ultracentrifuge pellet of M13 phage? I'd guess this is a CsCl cushion method . . . Thanks Daniel Kim dkim@nmsu.edu From owner-repertoires@net.bio.net Tue Oct 29 22:00:00 1996 Path: biosci!novell.pg.tno.nl!R.HANEMAAIJER From: R.HANEMAAIJER@novell.pg.tno.nl ("R Hanemaaijer") Newsgroups: bionet.molecules.repertoires Subject: FAb-isolation\purification Date: 30 Oct 1996 02:57:42 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 17 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net I've a question about the isolation\purification of Fab's. We have selected specific Fab's from a human library and isolated them from the E. Coli's by the freeze-dry method. We would like to use them as catching antibodies coated on 96-well plates and therefore need to purify them. Has anyone of you good experiences with the purification of Fab's ? We tried protein G (Pharmacia claimed that protein G binds both intact antibodies and Fabs), but the few Fab's we tried with protein G did not bind. We cannot use a ligand-coated column, because the amount of protein is limited. Hope someone can help me, Thanks, Roeland Hanemaaijer From owner-repertoires@net.bio.net Tue Oct 29 22:00:00 1996 Path: biosci!daresbury!not-for-mail From: Franco Felici Newsgroups: bionet.molecules.repertoires Subject: Re: ultra pellet of M13 Date: 30 Oct 1996 15:50:35 -0000 Lines: 12 Sender: lpddist@mserv1.dl.ac.uk Distribution: bionet Message-ID: <557tgb$ec6@mserv1.dl.ac.uk> Original-To: molreps@dl.ac.uk Daniel Kim asks: >Can anyone tell me what are good conditions for doing an ultracentrifuge >pellet of M13 phage? Phage can be pelleted from aqueous suspensions through ultracentrifugation at 50,000 rpm for 4 hours at 4 C in a Beckman 70Ti rotor (or equivalent). Best wishes, Franco Felici felici@irbm.it