Flow-through peptide synthesis and cell-based assays on Teflon-coated paper
The synthesis of peptides and proteins is an extremely complex matter, because they must be built up one amino acid at a time in repeated reaction and washing steps. Solid-phase synthesis has made this easier. Canadian researchers introduced a new variant of this technique in the journal Angewandte Chemie.
In their system, parallel syntheses take place in small regions on a paper support. The resulting peptide arrangements can be used in cell-based assays or in the search for 3D materials that support cell adhesion and growth.
In classical solid-phase synthesis, the growing peptide chain is fixed to a solid support—usually a polymer bead—so that the reagents can be rapidly and easily washed away after each step. Parallel solid-phase synthesis, known as SPOT synthesis, was developed as an alternative in the 1990s. This allows a large number of peptides to be obtained on a planar support with a small surface area.
SPOT synthesis has since been adapted for other applications, such as cell-based screening. The problem is that existing SPOT systems are not well-suited for chemical reactions. When individual drops of reagent are added by pipette, they wet small areas of the membrane—the SPOTs.
The circular spot of solvent absorbed by the membrane determines the size of the “reaction vessel”. Unlike in classical solid-phase synthesis, this limits the amounts of reagent, and flow-through conditions are not possible. This significantly limits the possible yields of the reactions.
A team headed by Frédérique Deiss and Ratmir Derda at the University of Alberta (Canada) has now found an elegant solution to this problem. The researchers used a Teflon coating to form a pattern of solvent-repellent barriers on a paper support. The pattern restricts the liquids to specific Teflon-free zones on the paper, forming small “reaction vessels” that can hold a larger volume than the usual SPOTs.
This not only allows for the use of excess volumes of reagents, but also allows for a flow-through reaction because the larger volume ensures for gravity-driven flow of the reagent solution through the paper. The flow rate can be varied by using paper of different porosity. This significantly improves yields.
There is an additional advantage to this method: the paper can be stacked or folded into thicker three-dimensional structures. The researchers were able to identify various peptides among those immobilized on the surface that support cell adhesion, growth, or differentiation in a three-dimensional environment.
About the Author
Ratmir Derda started his career as an assistant professor at the department of Chemistry at the University of Alberta in 2011. He is a principal investigator at the Alberta Glycomics Centre and Sentinel Bioactive Paper Network. In 2012, he received a Rising Star in Global Health Award from Grand Challenges Canada.
Author: Ratmir Derda, University of Alberta, Edmonton (Canada), http://derda.chem.ualberta.ca/contact/
Title: Flow-Through Synthesis on Teflon-Patterned Paper to Produce Peptide Arrays That Can Be Used for Cell-Based Assays
Angewandte Chemie International Edition, Permalink to the article: http://dx.doi.org/10.1002/anie.201402037
Ratmir Derda | Angewandte Chemie
Unexpected flexibility found in odorant molecules
27.06.2016 | Max-Planck-Institut für Struktur und Dynamik der Materie
Newly-discovered signal in the cell sets protein pathways to mitochondria
27.06.2016 | Eberhard Karls Universität Tübingen
High resolution rotational spectroscopy reveals an unprecedented number of conformations of an odorant molecule – a new world record!
In a recent publication in the journal Physical Chemistry Chemical Physics, researchers from the Max Planck Institute for the Structure and Dynamics of Matter...
Strands of cow cartilage substitute for ink in a 3D bioprinting process that may one day create cartilage patches for worn out joints, according to a team of engineers. "Our goal is to create tissue that can be used to replace large amounts of worn out tissue or design patches," said Ibrahim T. Ozbolat, associate professor of engineering science and mechanics. "Those who have osteoarthritis in their joints suffer a lot. We need a new alternative treatment for this."
Cartilage is a good tissue to target for scale-up bioprinting because it is made up of only one cell type and has no blood vessels within the tissue. It is...
Physicists in Innsbruck have realized the first quantum simulation of lattice gauge theories, building a bridge between high-energy theory and atomic physics. In the journal Nature, Rainer Blatt‘s and Peter Zoller’s research teams describe how they simulated the creation of elementary particle pairs out of the vacuum by using a quantum computer.
Elementary particles are the fundamental buildings blocks of matter, and their properties are described by the Standard Model of particle physics. The...
A year and a half on the outer wall of the International Space Station ISS in altitude of 400 kilometers is a real challenge. Whether a primordial bacterium...
Researchers at Case Western Reserve University have developed a way to swiftly and precisely control electron spins at room temperature.
09.06.2016 | Event News
24.05.2016 | Event News
20.05.2016 | Event News
27.06.2016 | Agricultural and Forestry Science
27.06.2016 | Physics and Astronomy
27.06.2016 | Life Sciences