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Chromatography & electrophoresis
glossary & taxonomy
Evolving terminologies for emerging
technologies
Suggestions? Comments? Questions? Mary Chitty MSLS mchitty@healthtech.com
Last revised
September 18, 2018
Technologies term index Related glossaries include Bioprocessing Mass
Spectrometry
Proteins
Proteomics Sequencing
2D See Two D affinity chromatography:
A selective separation technique by which a
compound (e.g., an antibody) is immobilized on a polymeric matrix and
used to bind selectively other compounds. Following removal of the unattached
components, the bound compound is displaced by changing the concentration of
protons, salts, or cofactors in the eluent. IUPAC Biotech
CE-MS (Capillary Electrophoresis- Mass Spectrometry): Separation
is achieved through channels etched on the surface of the capillary (connected
to an external high- voltage power supply) which delivers sample to ESI- MS.
Automatable approach, with great sensitivity.
CIEF Capillary electrophoresis IsoElectric Focusing:
Sample is “focused” in the capillary tube, both separating and concentrating
the protein or peptide at its isoelectric point. Then the entire mixture
is delivered to the mass spectrometer. Using this technique the researchers
have been able to record the molecular weight of all the proteins under
study. Can be used to separate proteins from microorganisms.
capillary array:
A bundle of extremely narrow gel- filled tubes
used to significantly speed the process of separating biological material.
See capillary electrophoresis [ALSSA]
capillary
electromigration: Capillary
electromigration techniques are becoming increasingly popular and important in
chemical analysis, especially in the bioanalytical field. ... The separations in
capillary electromigration techniques are achieved in narrow capillaries by
employing a high electric field strength. These techniques include capillary
electrophoretic techniques and electrically driven capillary chromatographic
techniques, based on different separation principles. In some cases, these
principles overlap. Capillary electromigration techniques have proved to be
highly effective for the separation of small organic and inorganic ions,
pharmaceuticals, explosives, dyes, polymers, proteins and peptides, DNA and RNA,
cells, particles, etc. [5–10] IUPAC Recommendations,
Terminology for Analytical Capillary Electromigration Techniques, 2003 http://www.iupac.org/publications/pac/2004/pdf/7602x0443.pdf
Capillary Electrophoresis CE: A very sensitive separation
technology, uses a capillary tube. Narrower terms: capillary gel electrophoresis, microcapillary
electrophoresis
capillary gel electrophoresis:
Gel is in a capillary microchannel. Improved quantification for
highly complex samples. Related term: microcapillary electrophoresis.
carrier
ampholyte(s):
Small soluble molecules that are capable
of acting as an acid or a base in standard ISO-DALT 2D
gel electrophoresis.
chromatography: A physical method of separation in which the components to be separated are distributed between two phases, one of which is stationary (stationary phase) while the other (the mobile phase) moves in a definite direction.
IUPAC Compendium Techniques used to separate mixtures of substances based on differences in
the relative affinities of the substances for mobile and stationary phases. A
mobile phase (fluid or gas) passes through a column containing a stationary
phase of porous solid or liquid coated on a solid support. Usage is both
analytical for small amounts and preparative for bulk amounts. MeSH, 1966
DGGE (Denaturing Gradient Gel Electrophoresis): A technique for
screening for SNPs. dHPLC
denaturing high- performance
liquid chromatography:
Used in SNP- detection methods based on
discrimination between perfect and mismatched hybridization, measures melting
temperatures. (Mismatching results in lowered melting temperature.) This method
involves passing the hybridization mixture through a chromatographic column
several times at different temperatures and deriving the melting temperature
from changes in the chromatogram. This method has the advantage of not needing
labeled components; however, each assay requires at least 15 minutes to
complete.
dielectrophoresis: http://en.wikipedia.org/wiki/Dielectrophoresis
Edman degradation:
A lab technique used to find out the order
of amino acids in a polypeptide (chain of amino acids). It involves using
the Edman reagent, phenyl isothiocyanate (PITC), to react one by one with
each amino acid, in order. The technique is used in machines which automatically
sequence (determine the order of subunits) polypeptides. [OMD]
electrophoresis: The motion of colloidal particles in an
electric field. IUPAC Compendium
Technique for separating and purifying molecules according to the relative
distance they travel under the influence of an electric current. [NHLBI]
An electrochemical process in which macromolecules or colloidal particles
with a net electric charge migrate in a solution under the influence of an
electric current. MeSH
A method of separating large molecules (such as DNA fragments or proteins)
from a mixture of similar molecules. An electric current is passed through
a medium containing the mixture, and each kind of molecule travels through
the medium at a different rate, depending on its electrical charge and
size. Separation is based on these differences. Agarose and acrylamide
gels are the media commonly used for electrophoresis of proteins and nucleic
acids. [DOE]
A technique for separating and analyzing substances based
on the movement of their ions in an electrical field. Related term: gel
electrophoresis
Expanded Bed Adsorption
EBA:
What
chromatographers need is a method that combines sample preparation with
the first stage of chromatography. Welcome to expanded- bed adsorption
(EBA)
chromatography. EBA uses apparatus that is familiar to most users of
standard liquid chromatography. The column has a flow adapter that is positioned
to suit the specific step of resin preparation or protein purification. And a
series of pumps and valves, connected through the adapter and bottom of the
column, control the flow rate and direction of the buffer and sample loading (1,
2). Thus, it is feasible to perform preliminary EBA trials with a
little ingenuity and standard chromatographic equipment. ...In
the more traditional packed-bed methods, where the resin is confined between the
bottom of the column and the flow adapter, clogging occurs when particulate
matter and cell debris cannot flow around the closely packed resin beads. In
contrast, EBA columns are fed from below, and the adapter is held away from the
packed resin level, giving the resin room to expand and thus creating spaces
between the beads. Randall C. Willis "Expanded Bed Adsorption" Modern
Drug Discovery 4 (12): 43- 44 Dec. 2001 http://pubs.acs.org/subscribe/journals/mdd/v04/i12/html/12toolbox.html
frontal affinity chromatography:
Method for screening mixtures
of compounds for affinity against an immobilised target. IUPAC COMBINATORIAL
CHEMISTRY
gel electrophoresis:
A DNA separation technique that is very
important in DNA sequencing. Standard sequencing procedures involve cloning
DNA fragments into special sequencing cloning vectors that carry tiny pieces
of DNA. The next step is to determine the base sequence of the tiny fragments
by a special procedure that generates a series of even tinier DNA fragments
that differ in size by only one base. These nested fragments are separated
by gel electrophoresis, in which the DNA pieces are added to a gelatinous
solution, allowing the fragments to work their way down through the gel.
Smaller pieces move faster and will reach the bottom first. Movement through
the gel is hastened by applying an electrical field to the gel. [DOE]
Electrophoresis performed in a polymer matrix (gel) Broader term:
electrophoresis,
PAGE, SDS- PAGE Narrower terms: 2D gel electrophoresis, pulsed- field gel
electrophoresis
gel shift:
A method by which the interaction of a nucleic acid (DNA or RNA) with a protein is detected. The mobility of the nucleic acid is monitored in an agarose gel in the presence and absence of the protein: if the protein binds to the nucleic acid, the complex migrates more slowly in the gel (hence "gel shift"). A
"supershift" allows determination of the specific protein, by virtue of a second shift in mobility that accompanies binding of a specific antibody to the nucleic acid-protein complex.
Lemon, Stanley M. and Alan Barbour "Glossary of terms commonly used in
molecular biology" UNC- Chapel Hill and Univ. of Texas Health Science
Center, US, 2000 http://www.med.unc.edu/wrkunits/3ctrpgm/pmbb/mbt/GLOS.htm
glass capillary electrophoresis:
. A type of automated
sequencer based on glass capillary electrophoresis. Each automated sequencer
possesses an array of capillaries that contain the DNA separation matrix.
Fluorescently labeled DNA is applied by electrokinetic injection into each
capillary, resolved by electrophoresis through the capillary matrix, and then
detected as it passes by a clear window in each capillary or as the DNA
fragments exit the end of each capillary. Before the next run, the instrument
completely recycles itself, injecting new separation matrix into each capillary
and preparing the instrument for data collection.
HPLC High Performance Liquid Chromatography:
Liquid chromatographic
techniques which feature high inlet pressures, high sensitivity, and high
speed. MeSH, 1981
HPLC
Center Glossary of terms, Upchurch Scientific, http://webstore.idex-hs.com/TechInfo/glossary.asp
IEF (IsoElectric Focusing).
Electrophoresis in which a pH gradient is
established in a gel medium and proteins migrate until they reach the site (or
focus) at which the pH is equal to their isoelectric point. MeSH, 1974 First reported in 1961. Earliest version used carrier ampholytes;
newer uses immobilized pH gradients. Compare with yeast two hybrid.
Proteomics
IPG (Immobilized pH Gradient).
Now standard 2D gel electrophoresis
method. Amersham Pharmacia led the commercialization effort. Can be used
for pH range 3-12 and is method of choice for IsoElectric Focusing. Narrower term:
immobilized pH gradient shift.
IPG-DALT:
Immobilized pH Gradient with protein mass expressed
in Daltons.
ISO-DALT:
ISOelectric focusing electrophoresis, measured in Daltons.
Cannot resolve basic proteins with pI > 7.0-7.5 pH. Use of isoelectric
focusing greatly improves resolution.
immobilized pH gradient shift: Introduction of gels with immobilized
pH gradient (in 1982) improved the separability of 2D gel electrophoresis. Narrower term:
IPG.
immobilized pH gradient strips, very narrow range:
Also called
“zoom gels” or proteomic contigs, used to improve sensitivity of 2D gel
electrophoresis. Broader term IPG.
immunoprecipitation:
Antibody based affinity purification.
in gel protein digestion:
A method to cleave proteins into more
easily identifiable peptide subunits directly in the gel immediately following
separation of the proteins by gel electrophoresis. This method uses different
enzymes, which specifically cleave between certain amino acid sequences. ALSSA isoelectric point:
The pH in solutions of proteins and related
compounds at which the dipolar ions are at a maximum. MeSH, 1991
The pH value at which the net electric charge of an
elementary entity is zero. pI is a commonly used symbol for this kind- of-
quantity. It should be replaced by pH (I) because it is a pH determined under
that particular condition. IUPAC Compendium liquid chromatography LC:
A separation technique in which the
mobile phase is a liquid. Liquid chromatography can be carried out either
in a column or on a plane. Present- day liquid chromatography generally
utilizing very small particles and a relatively high inlet pressure is
often characterized by the term high- performance (or high- pressure)
liquid chromatography, and the acronym HPLC. IUPAC Compendium
Chromatographic techniques in which the mobile phase is a liquid.
MeSH, 1976
microcapillary electrophoresis (micro-capillary electrophoresis):
Is this different from microchip electrophoresis?
microscale HPLC systems: Nanoscience
& miniaturization glossary
multidimensional
chromatography: The use of two or more columns or
chromatographic techniques which enable a better separation. It is useful for
sample cleanup, increased resolution, and increased throughput. It can be used
off- line by collecting fractions and reinjecting onto a second column or on-
line by the use of a switching valve. It also called coupled column
chromatography, column switching, multicolumn chromatography, or boxcar
chromatography. HPLC Glossary, Waters Corp.,
2003 http://www.waters.com/watersdivision/images/aboutus/hplcglossary.htm#M
NEPHGE: Non-Equilibrium PH Gradient Electrophoresis. Nonequilibrium
pH gel electrophoresis (NEPHGE) is a technique developed to resolve proteins
with extremely basic isoelectric points (pH 7.5-11.0) (1,2).
These proteins are difficult to resolve using standard IEF, because the presence
of urea in IEF gels has a buffering effect and prevents the pH gradient from
reaching the very basic values (with a pH above 7.3-7.6) (3). In addition,
cathodic drift causes many very basic proteins to run off the end of the gel.
Mary Lopez The Protein Protocols Handbook
2002, Part II, ,
DOI: 10.1385/1-59259-169-8:181
http://www.springerlink.com/content/q6vp3t06vx6648r1/
non-denaturing gel electrophoresis:
DNA fragments separate in
a double stranded form. Also known as “native gel” systems.
Related term: IsoElectric Focusing.
pI: See isoelectric point.
PAGE PolyAcrylamide Gel Electrophoresis:
Electrophoresis
in which a polyacrylamide gel is used as the diffusion medium. [Metathesaurus]
Narrower term: SDS-PAGE.
proteomic contigs: See immobilized pH gradient strip, very narrow range.
pulsed field gel electrophoresis:
Electrophoresis in which the
direction of the electric field is changed periodically. This technique is
similar to other electrophoretic methods normally used to separate double-
stranded DNA molecules ranging in size up to tens of thousands of
base-pairs. However, by alternating the electric field direction one is able to
separate DNA molecules up to several million base- pairs in length. MeSH, 1992
SDS-PAGE (Sodium Dodecyl Sulfate PolyAcrylamide Gel Electrophoresis):
May be helped by mass spectrometry, can be blotted or eluted for further
chemical analysis i.e. Edman degradation, amino acid microsequencing or
scanned MALDI-MS. Also known as denaturing gel electrophoresis or native
gel electrophoresis. [Glick] Broader terms: gel electrophoresis, PAGE.
Supercritical Fluid Chromatography SFC:
A chromatography
method using supercritical fluid, usually carbon dioxide under very high pressure (around 73 atmospheres or 1070 psi at room temperature) as the mobile phase. Other solvents are sometimes added as modifiers. This is used both for analytical (SFC) and extraction (SFE)
purposes. MeSH, 2002
tandem chromatography:
Tandem chromatography is used to automatically collect selected fractions
from one column and inject them onto a second column. Eluted fractions from the
first column are collected into a DynaLoop or other sample loop. After injection
of the collected sample, it is eluted from the second column. ... This
automated procedure can be performed at neutral pH, as an alternative to
affinity chromatography, which requires low-pH buffers to elute the antibodies.
Basic Principles of Chromatography, Dept of Chemistry, Minnesota State
University Moorhead
http://www.mnstate.edu/biotech/FPLC_talk_hndout.pdf
Two D DIGE: 2D Differential (or Difference) In Gel Electrophoresis
Two D gel electrophoresis:
Electrophoresis in which a second perpendicular electrophoretic transport is
performed on the separate components resulting from the first electrophoresis.
This technique is usually performed on polyacrylamide gels. MeSH, 1989
Proteins are first separated
across a gel according to their isoelectric point, then separated in a
perpendicular direction on the basis of their molecular weight. First introduced
in 1975. Most commonly used method for protein separation in proteomics.
Two-Dimensional Difference Gel Electrophoresis:
Methods of comparing two or more samples
on the same two-dimensional gel electrophoresis gel. MeSH 2011
zoom gels:
See immobilized pH gradient strips, very narrow
range.
Chromatography & Electrophoresis resources
Glossary of HPLC Separation
Terms LCGC
Feb 01, 2008, By Peter W. Carr, Ronald E. Majors
LCGC North America, Volume 26, Issue 2, pg 118–168
http://www.chromatographyonline.com/glossary-hplclc-separation-terms?id=&pageID=1&sk=&date= HPLC
Center Glossary of terms, Upchurch Scientific, 2005,
about 250 terms http://www.upchurch.com/TechInfo/glossary.asp
IUPAC International Union of Pure and Applied Chemistry, Compendium of
Chemical Terminology: Recommendations, compiled by Alan D. McNaught and
Andrew Wilkinson, Blackwell Science, 1997. "Gold Book" 6500+
definitions. http://goldbook.iupac.org/
IUPAC International Union of Pure and Applied Chemistry, Glossary for
Chemists of terms used in biotechnology. Recommendations, Pure & Applied
Chemistry 64 (1): 143-168, 1992. 200 + definitions.
IUPAC
Terminology for Analytical Capillary Electromigration Techniques,
International Union of Pure and Applied Chemistry, 2003, 24 definitions and 40+
acronyms and symbols. http://www.iupac.org/publications/pac/2004/pdf/7602x0443.pdf
IUPAC Chromatography
International Union of Pure and Applied Chemistry, Nomenclature for chromatography,
L.S. Ettre, editor, Pure and Applied Chemistry 65: 819-872, 1993.
450+ definitions http://www.iupac.org/publications/pac/1993/pdf/6504x0819.pdf SeparationsNow, Electrophoresis, Wiley
http://www.separationsnow.com/electrophoresis?tzcheck=1
SeparationsNow, Gas Chromatography, Wiley
http://www.separationsnow.com/gc
SeparationsNow, HPLC, Wiley
http://www.separationsnow.com/hplc
Wikipedia, Chromatography terms http://en.wikipedia.org/wiki/Chromatography#Chromatography_terms
15 definitions
IUPAC definitions are reprinted with the permission of the International
Union of Pure and Applied Chemistry.
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