PepArray™ Technology for Proteomics Applications
Custom high density peptide synthesis and multiplex proteomics assays on a microchip based on the µParaflo® microfluidics technology.
PepArray™ technology involves in situ (on chip) high density peptide synthesis and multiplex protein assays carried out in a microfluidic picoliter scale microarray. The PepArray™ is an assay reaction device which allows thousands of titration reactions, multiple step enzymatic reactions, and other biochemical reactions normally carried out in individual micro tubes or microtiter plates to be performed simultaneously in a single experiment. Our PepArray™ products and services are powerful tools for diverse proteomics applications, including: enzymatic assays for protein kinases and their inhibitors, epitope mapping, antibody and autoantibody detection, and proteomic profiling of phosphoprotein-binding proteins and domain-binding proteins.
The technology allows us to synthesize thousands of custom peptides (sequences can be defined to each single amino acid residue) on an addressable array to act as kinase substrates, antibody epitopes, or protein binding ligands. We can perform enzymatic/binding reactions in a high-throughput format and generate quantitative results in a controllable, enclosed environment with minimal sample usage. The assay results are analyzed by a microarray imager and standard microarray analysis software.
PepArray™ technology is made available to you as a service for multiple applications. Send us your sample(s) and we will perform the experiments you desire in our lab and send you back a detailed data report.
Microfluidic PepArray™ Design
Our array platform is unique and was developed based on proprietary microfluidics technology. We take advantage of microfluidics during both the synthesis of the arrays and during the binding/enzymatic assay reactions.
Synthesis – Our technology enables us to perform custom synthesis of peptides and peptidomimetics directly on the high density microfluidic chip at addressable chip locations. We synthesize all peptide microarrays to order using either a catalog design or a design generated from a list of peptides provided by customer or compiled from peptide databases. We do not need to pre-synthesize and then spot the arrays. Therefore, there is no requirement for significant up-front investment in synthesis of peptide libraries and we do not have a stock-pile of arrays sitting on a shelf with content that may become out of date. We have array formats of 4K, 5K, and 30K features.
Assays – The chip is an enclosed microfluidic system, consisting of fluid distribution channels and picoliter scale reaction chambers. The chip is a closed system, unlike an open glass slide as with typical microarrays so dye oxidation and deterioration are not an issue. Miniaturized microfluidic array features reduce consumption of samples to the sub-nanoliter to picoliter level per reaction. To perform the binding/enzymatic reactions, the solution is re-circulated through the chip under defined environmental conditions, time, temp, flow rate, and direction, not merely pipetted on top of the array. Active circulation of the sample throughout the chip produces a uniform distribution of the sample solution on the array that enhances the binding kinetics and stringency wash processes. This results in increased signal-to-noise and decreased CV values. There are no “dark” areas due to non-uniform sample distribution. Additionally, microfluidic PepArrays™ can be used for repetitive time-, protein/antibody/enzyme concentration-, or co-factor/inhibitor-dependent measurements. The array can be used multiple times, each time assay is carried out and assay signals are collected by fluorescence scan. These assays generate a very large set of data for proteomic assays and valuable information which would take weeks and months to produce using a conventional single protein/antibody/enzyme assay kit.
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PepArray™ platform technology offers in situ synthesis of peptides and peptidomimetics using conventional peptide chemistry in an addressable format and offers the capability for massively parallel quantitative measurements at a scale that was previously not available. It is a flexible platform which can be engineered to adapt to many different applications. This means high throughput peptide drug discovery screening can be carried out on a comprehensive scale and in a systematic way. Not only is this technology enabling therapeutic synthetic peptide screening through rapid design, synthesis, and screening of diverse peptides and peptide analogs against important therapeutic targets, but it is also enabling the use of peptides as tools for study of kinases, antibodies and auto-antibodies, phosphopeptide-binding proteins, and other pathway signaling molecules.
Perform protein kinase reactions on arrays of comprehensive sets of substrates. Measure kinase activity and inhibitor effects. Obtain in parallel hundreds of reaction profiles and curves for parameters such as VMAX, KM, and Ki. Use of microarray titer plate format will increase your workflow by at least 40-fold that of a single microtiter plate.
Perform large scale parallel epitope mapping from pathogens. Identify high affinity antibody binding peptides. Screen sensitive antibody-detection peptides. Detect autoantibody/biomarker from serum or biological samples, determine Kd (binding affinity) of hundreds of binding peptides, obtain titration curves for antibodies.
Profile SH2-containing proteins in complex biological samples by an array of phosphopeptides which are carefully curated from literature. Screen effective SH2 binding inhibitors and measure inhibitory activities. Screen signature phosphopeptides for protein capture as a tool for signaling pathway characterization.
Screen biomarkers (proteins and peptide binding targets) from biological samples such as blood serum or cell lysates. Use novel protein capture peptides to detect and profile on the same chip platform a wide range of molecular biomarkers: signaling protein kinases, pathway phosphoprotein-binding and domain-binding proteins, immuno-responsive antibodies, metabolites and metabolic molecules, and other small molecules.
Therapeutic Synthetic Peptide Screening
Rapidly design, synthesize, and screen diverse peptides, peptide analogs, and peptides modified with glyco-units against important therapeutic targets.
- Custom synthesis – in situ (on chip) high density peptide synthesis means arrays are made to order on an addressable array (sequences can be defined to each single amino acid residue).
- No spotting – we do not need to pre-synthesize and then spot the arrays so there is no up-front investment in synthesis of peptide libraries.
- Always up-to-date – no stock-pile of arrays sitting on a shelf with content that may become out of date.
- Microfluidics – array platform is unique and was developed based on a powerful proprietary microfluidics technology.
- High-throughput – the PepArray™ is an assay reaction device which allows thousands of proteomics reactions at once normally carried out in individual micro tubes or microtiter plates.
- Picoliter scale reactions – high-throughput format generates quantitative results with minimal sample usage (sub-nanoliter to picoliter level per reaction).
- Closed system – the chip is a closed system, not an open glass slide so dye oxidation and deterioration are not an issue.
- Controlled environment – the reaction solution is re-circulated through the chip under defined environmental conditions, time, temp, flow rate, and direction, not merely pipetted on top of the array.
- Enhanced binding kinetics – active circulation of the sample throughout the chip produces a uniform distribution of the sample solution on the array that enhances the binding kinetics and stringency wash processes. This results in increased signal-to-noise and decreased CV values.
- No “dark” areas – due to non-uniform sample distribution.
- Flexible format – we have array formats of 4K, 5K, and 30K features, suitable for diverse applications.
- Comprehensive service – PepArray™ technology is made available to you as a comprehensive service for many applications.
- Diverse proteomics applications – enzymatic assays for protein kinases and their inhibitors, epitope mapping, antibody and autoantibody detection, and proteomic profiling of phosphoprotein-binding proteins and domain-binding proteins.