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Klas Ola Blixt answers a few questions about this month's
new hot paper in the field of Biology & Biochemistry.
From
•>>May 2006
Field:
Biology & Biochemistry
Article Title: Printed covalent glycan array for ligand profiling of diverse glycan binding proteins
Authors: Blixt,
O;Head, S;Mondala, T;Scanlan, C;Huflejt,
ME;Alvarez, R;Bryan, MC;Fazio, F;Calarese, D;Stevens, J;Razi,
N;Stevens, DJ;Skehel, JJ;van Die, I;Burton, DR;Wilson,
IA;Cummings, R;Bovin, N;Wong, CH;Paulson, JC
Journal: PROC NAT ACAD SCI USA
Volume: 101
Issue: 49
Page: 17033-17038
Year: DEC 7 2004
* Scripps Res Inst, Glycan Synth & Prot Express Core D, Consortium Funct
Glycom, 10550 N Torrey Pines Rd, La Jolla, CA 92037 USA.
* Scripps Res Inst, Glycan Synth & Prot Express Core D, Consortium Funct
Glycom, La Jolla, CA 92037 USA.
* Scripps Res Inst, DNA Microarray Core Facil, La Jolla, CA 92037 USA.
* Scripps Res Inst, Dept Mol Biol, La Jolla, CA 92037 USA.
* Scripps Res Inst, Dept Chem, La Jolla, CA 92037 USA.
* Scripps Res Inst, Dept Immunol, La Jolla, CA 92037 USA.
* Sidney Kimmel Canc Ctr, San Diego, CA 92121 USA.
* Univ Oklahoma, Hlth Sci Ctr, Dept Biochem & Mol
Biol, Oklahoma City, OK 73104 USA.
* Natl Inst Med Res, Med Res Ctr, London NW7, England.
* Vrije Univ Amsterdam, Ctr Med, Dept Mol Cell Biol &
Immunol, NL-1007 MB Amsterdam, Netherlands.
* Russian Acad Sci, Shemyakin & Ovchinnikov Inst Bioorgan
Chem, Moscow 117997, Russia.
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 Why
do you think your paper is highly cited?
Our paper describes the first example of a highly diverse
oligosaccharide microarray. From this work, a significant
breakthrough comes about from the cumulative participation of a
number of synthetic carbohydrate chemists to provide a unique
diversity within the 300-member array, which clearly offers a unique
advantage in favor of the Consortium for Functional Glycomics (CFG).
 View
image of the array technology.
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“This field is a necessary component in
understanding human health because of the importance of glycans as key players in all the
body's functions.”
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Amino-functionalized glycans are covalently attached through
robotic imprinting onto commercially available N-hydroxysuccinimide
activated glass slides, a method that is firmly established for
protein array technology. A second advancement in this work is the
creation of multivalent glycan-binding protein complexes through the
use of secondary antibodies to overcome low affinity. In addition, a
very low background binding with this platform enables analysis of a
broad spectra of GBP such as, mammalian lectins, antibodies, human
serum, intact virus, etc.
Does
it describe a new discovery or a new methodology that's useful to
others?
Our paper describes a breakthrough screening technology for
discovery in the field of glycomics. It has set the precedence
standard for the Consortium’s worldwide efforts to identify
biomedically important carbohydrate recognition specificities. The
Consortium offers free glycan-array-screening service to the
community, and the accumulated data are being updated into a central
database, which is also open to public access.
Could
you summarize the significance of your paper in layman's terms?
Many proteins involved in communication between cells recognize
glycan structures on cell surfaces. Glycans also have a pivotal
function in diverse areas of biology and medicine, including
infectious diseases, cancer progression, inflammation, asthma,
Alzheimer’s disease, nerve and neuromuscular diseases, and many
others.
Our glycan array technology is a glass slide, onto which hundreds
of different glycan chains are printed. The array offers scientists
a cutting-edge research tool allowing them to analyze the
specificities of GBPs, which function through their binding to such
sugar chains.
The functional glycan microarray will speed research in glycomics,
because it will allow scientists to determine to which carbohydrate
structures these proteins bind. The microarray will transform
research in the burgeoning field of glycomics, which is devoted to
the systematic identification and characterization of all the glycan
structures produced by organisms. This field is a necessary
component in understanding human health because of the importance of
glycans as key players in all the body’s functions.
How
did you become involved in this research, and were any problems
encountered along the way?
During the initial funding period of CFG (2001- ), the Glycan
Array Synthesis Core and Glycanprotein Interaction Core facilities
have been devoted to the development of tools and resources for the
field. To jump-start the glycan array effort, a platform based on a
96-well microtiter plate format using biotinylated glycans was
evaluated.
During the first years, the microtiter plate array assay was used
extensively to assay GBPs for glycan-binding specificity. The major
strength of the plate arrays was the large number of glycans with
diverse structures that can be tested as ligands for GBPs. The two
limitations of the plate array were the amount of GBP required for
an assay (800-1000 ug) and the need for multiple plates per GBP as
the glycan array continues to expand.
Even with these two limitations, the plate array screening
service provided by CFG could not be duplicated by any other
academic or commercial facility. The printed glycan arrays that were
subsequently developed successfully addressed the two limitations of
the plate array and have become the primary workhorse of CFG. An
additional advantage of the printed glycan array is the presence of
multiple spots of each glycan on the slide, thereby allowing
statistical analysis of glycan binding data.
Ola Blixt, Ph.D.
Assistant Professor
Core Director
Glycan Array Synthesis Core-D Consortium for Functional Glycomics
The Scripps Research Institute
Department of Molecular Biology
La Jolla, CA, USA
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ESI Special Topics,
May 2006
Citing URL - http://www.esi-topics.com/nhp/2006/may-06-Blixt.html
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