Isoelfocus 3.0 (*)
A multimedia computer-aided learning program on Isoelectric Focusing
and its derived techniques as tools in biomedecine and agrofood industries.
( IDDN.FR.001.510016.R.C.1997.000.31235 )
PC and MAC version
Screen display of oral commentary.
Optional connexion to Web sites.
Authors : ** CADE-TREYER Denise; CADE Ambroise ; ***DARJO
Alain ; JOUVION-MORENO Myrian
University Pierre & Marie CURIE 4, Place Jussieu 75 252 PARIS Cedex 05
**e-mail : cade.denise@wanadoo.fr
***e-mail : alain.darjo@wanadoo.fr
(*)For users of the preceding version 1.0, see the
remark on the new downloading process in 6.Updating.
Isoelectric focusing (IEF) is the high resolution electrophoretic separation, in a pH gradient of charged amphoteric molecules, notably proteins. With its developments and derived techniques (2D-electrophoresis, Capillary IEF...) IEF has gained increasing importance, as a tool in many, biomedical and agrofood research and industrial fields. There is therefore a high demand for thorough theorical and practical understanding of the basics of IEF, not only for students’education, but also, for scientists, clinicians, engineers and technicians.
«Isoelfocus» is a multimedia interactive computer-aided learning
program, which should be a stimulating approach of IEF, before to dive into more refined
and exhaustive printed textbooks on the subject. With easy updating and customizing, it
lends itself well to teaching.
At the heart of IEF methods lies the titration curve, which is the display of the charge
behaviour, along a pH gradient, of a protein to be separated. The titration curve of each
protein predicts its isoelectric point, which is the pH where the net charge of the
protein is zero. Isoelfocus has thus implemented the titration curve as a comprehensive
tool for IEF-based methods.
Some relevant examples will highlight the nowadays wide-spread use of IEF technologies :
 | Hospital reference centers of drepanocytocis use IEF as a routine test for diagnosis of sickle-cell anemia, the most prevalent genetic disease in African regions rife with malaria (1% prevalence) and in some european or american highly urbanized regions with black ethnic groups (1/500 births). |
| IEF of glycosylated hemoglobin, a reliable indicator of blood sugar level, is currently used in clinical monitoring of diabetics. |
| In clinical genetics, IEF phenotyping of blood proteins (PGM1, transferrin etc.) is commonly used. |
| In biotechnology and pharmaceutical industries, IEF fingerprinting of monoclonal or recombinant antibodies is required for database registration or for release assays of diagnostic kits or of glycoforms of recombinant proteins as therapeutic products. Recent optimization has allowed capillary IEF to be validated, for these purposes, as an automated quantitative test. |
| In agrofood industries, IEF is one of the assays listed in FDA’s points to consider for protein quality control of foodstuffs and fraud surveillance for meat, fish, cheese, flour etc...from fresh to deepfrozen or canned products. |
| IEF tests are widely used for determination of cereal or vegetable varieties in agriculture or food processing industries, as required by law of most countries. At the international level, the CODEX (*) will doubtless be brought to recommend IEF for protein quality control and for variety determination. |
| Proteome research : 2D electrophoresis (IPG-dalt) has become the
reproducible separation method of reference, coupled with mass-spectrometry
or other analytical approaches and with web browser-based bioinformatic tools,
for protein mapping, sequencing, database development and screening, taxonomic studies,
molecular imaging and functional analysis, search for clinical markers as well as
for drug target design. (*) The CODEX (or Codex Alimentarius) is the international agency created by FAO and WHO in 1962 to harmonize the legislation in food exchanges.WTO (the World Trade Organization) must comply with the standards recommended by the CODEX to give a ruling on matters of contention in agrofood exchanges between countries. |
PC : 90 Mhz or faster PC ; at
least 8Mb RAM ; hard disk : 50 Mb free for loading the files ; Monitor : configured to VGA
(640 x 480) with 65000 colors ; sound card and speakers.
Windows 95 or later version.
MAC : hard disk : 50 Mb free for loading the files
; minimal multimedia configuration.
3. How to download and
install isoelfocus :
Isoelfocus is distributed free of charge.
Commercial use is not authorized. Users are under their own responsibility.
Isoelfocus is subdivided into several files for easy transfer.
Notice : in order to obtain an efficient transfer MS Internet Explorer should be used.
| Preparing PC version |
Then create a folder (named for instance "Isoelfocus"), at the place of your choice, in which the self extracting basePC.exe will be decompressed.
Then click on the basePC.exe file in C:\temp to install its content into the created folder.
| Preparing MAC version |
You have to download the following files : !Cthbsea.bin and !Aapisea.bin. Then you must expand these files (with Stuffit Expander). The resulting video files (Cthb1.avi and Api256.avi) must be stored into the folder you have created ("Isoelfocus"). Notice that despite their name they have the MOV format. These names must be kept unchanged.
If necessary recent versions of Stuffit Expander can be downloaded at :
http://www.stuffit.com/expander/download.html
| Downloading the Isoelfocus Modules on PC and MAC |
The modules are encoded in files suitable for PC and MAC.
Notice that each file can be downloaded and installed separately with time and run independently. The module 11 has been subdivided into 3 files.
Click on each file you want to download on the list of modules below. You must save each of them into the folder you have just created ("Isoelfocus" for instance).
| List of Modules | File | Ko | Update | |
| Module 1 | Overview of the applications of IEF | avprog1 | 4768 | 19/07/01 |
| Module 2 | The IEF Process | prog001 | 2555 | 19/07/01 |
| Module 3 | The titration curve of a single protein | prog003 | 1009 | 19/07/01 |
| Module 4 | The titration curve of a protein mixture | prog004 | 1540 | 19/07/01 |
| Module 5 | The titration curve as a tool for protein purification | postpr04 | 3871 | 19/07/01 |
| Module 6 | The resolving power in IEF | resolut | 1993 | 19/07/01 |
| Module 7 | The electric parameters | prog007 | 2302 | 19/07/01 |
| Module 8 | Generation of the pH gradient | gradient | 2250 | 19/07/01 |
| Module 9 | The pH gradient drift | gradrift | 955 | 19/07/01 |
| Module 10 | Immobilized pH gradients (IPG) | immobil | 3070 | 19/07/01 |
| Module 11 | Preparative IEF (1) | prepar | 547 | 19/07/01 |
| Module 11 | Preparative IEF (2) | prepar2 | 4418 | 19/07/01 |
| Module 11 | Preparative IEF (3) | prepar3 | 767 | 19/07/01 |
| Module 12 | 2D-Electrophoresis | bidim | 2516 | 19/07/01 |
| Module 13 | Capillary IEF | capill | 1906 | 19/07/019 |
| Module 14 | IEF Workshop | workshop | 631 | 19/07/01 |
| Introduction - Preface | intrelec | 2114 | 19/07/01 | |
| Main Menu - Key Words - References | routeur | 164 | 19/07/01 | |
| PC | Runtime Attached Files (Video, Xtras...) |
BASEPC.EXE | 4309 | 07/06/99 |
| MAC | Runtime Attached Files (Xtras) |
runa4m.hqx | 9076 | 04/08/97 |
Isoelfocus is displayed in a 640 X 480 window. So, for the best screen display, 640 X 480 resolution should be maintained.
First open the folder in which you have installed Isoelfocus.
On the list of directories and files displayed, it is mandatory to first double-click on the Runtime file (Runa4w32 for PC or Runa4m for MAC). A dialog box will then open and display the list of files (each file contains a module). On clicking on one of them you will run it.
Click on intrelec file to run once isoelfocus from the beginning (with introduction and preface).
Routinely, on clicking on the routeur file you will display the main menu, wherefrom each module can be run. The main menu gives also access to the keyword index and to the references.
Comments are welcome and may be addressed to the authors by e-mail : denc@ccr.jussieu.fr
Isoelfocus is on the Web since march 1998. Thanks to numerous suggestions or responses to RFC, the present version for PC and MAC has been improved and especially enriched with historical, research and technical comments or hyperlinks. The structure of Isoelfocus has been changed : the Runtime is from now on outside each module. Therefore users of the version 1.0 must download de novo the version 3.0 as a whole according to the process described in § 3. Users of the version 2.0 have only to download the files of the modules.
Further updates of the different modules will be displayed on this Web Page, on the table "list of modules" (column:update). You will have only to download an updated file to your Isoelfocus repertory. A revised module will automatically replace the previous version.
http://www.sfe-ices.org (French
Electrophoresis and Proteomic Analysis Society)
http://www.electrophoresis.org (Swiss
Electrophoresis Society)
http://www.harefield.nthames.nhs.uk/nhli/bes
(British Electrophoresis Society)
http://www.aesociety.org (American
Electrophoresis Society)
http://proteome.tmig.or.jp/JES/ (Japenese
Electrophoresis Society)
http://www.weihenstephan.de/blm/deg
(German Electrophoresis Society)
http://plab.ku.dk/tcbh/NoSSS.htm
(Nordic Separation Sciences Society)
http://www.aesociety.org/(Australian
Electrophoresis Society)
http://www.expasy.ch/ch2d/2DHunt (Expasy
Server with research tool for Web sites in Electrophoresis)
Isoelfocus was awarded the 1998 Amersham-Pharmacia Biotech Prize of Electrophoresis
at the Swiss and French Electrophoresis Societies joint Conference ( Basel 3 Dec 1998)
Isoelfocus obtained a Gold Award at the 1999 CERVOD (Concours d'Etudes et de Réalisations pour la Valorisation des Outils Didactiques) : French competition of studies and products designed to promote didactic tools.
