VirD™ Functional Membrane Protein Array

>400 Unique Proteins for Drug Development

VirD™ membrane protein arrays use a patented system to synthesize important drug targets with proper folding and function.

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Correctly-folded, Embedded, and Functional Membrane Proteins

VirD contains >400 unique proteins and is useful for drug development and functional assays

G protein-coupled receptors (GPCRs) play critical roles in a wide variety of physiological processes, and mutations in GPCRs are implicated in many diseases. GPCR receptors must be embedded in a cell membrane to fold correctly and are difficult to express in traditional recombinant expression systems. The most common method used to study GPCRs is to overexpress them in transfected mammalian cell lines. However, the transfected cells also contain endogenous receptors, making it a challenge to interpret experimental results.

Our VirD viral-based expression technology expresses individual GPCR proteins embedded in a cell membrane. Purified virions displaying correctly-folded GPCRs are printed onto glass to create VirD arrays. This method eliminates the problem of endogenous receptor competition when using traditional methods, and yields much cleaner data. VirD GPCR arrays can be purchased directly or provided as a complete analysis service.

VirD Membrane Protein Array Validation and Production

VirD is a microarray that uses virions to display functional membrane proteins like GPCRs and is useful for drug development and functional assays.

VirD Membrane Protein Array Validation and Production Figure

VirD Membrane Protein Array Methods and Analysis

VirD is a sensitive membrane protein matrix platform that provides quantitative binding interaction data from a few microliters of sample. VirD can help identify the function of orphan GPCRs and can be used to develop new drugs targeting membrane proteins and receptors.

VirD Membrane Protein Array Methods and Analysis Figure


VirD™ Service at CDI Labs

Technical Details

To create the VirD array, 500+ unique human membrane protein open reading frames (ORFs) are modified to remove their stop codons and subcloned into the UL27 locus of the herpes simplex (HSV-1) virus genome with a C-terminal v5 tag. These individually engineered viron clones are then used to infect mammalian cells. Native human protein assembly machinery correctly folds the proteins and shuttles them to the viral membrane. Membrane-protein bearing VirD virions are concentrated from culture supernatants and their membrane-protein expression confirmed via anti-V5 immunoblots.

Purified virions were then printed on SuperEpoxy glass slides to create the final GPCR-VirD arrays. The quality of printed VirD-GPCR arrays is confirmed via staining for HSV-1 Glycoprotein D (gD) via anti-gD antibody and comparing those as compared with bovine serum albumin negative controls (BSA).

Service Details and Data Deliverables

A simple antibody interaction VirD service involves drug candidate antibodies diluted and reacted with individual microarrays with BSA blocking. After incubation, samples are stained for one or two desired fluorescent secondary detection antibodies (i.e., IgG1). Arrays are scanned and raw TIF images are created using Molecular Devices Genepix microarray scanners. Spots are then aligned to CDI library (.gal) files, and spot readings are output as raw Genepix results (.gpr) files. These raw results are then delivered both as .gpr files and combined output data tables.

Sample Requirements

Antibody isolates10 μg per sample (concentration 0.1 mg/mL)
Protein / peptide / small molecule10 μg per sample (concentration 0.1 mg/mL)
RNA1 mL per sample (concentratiion 2 μM)
Other fluids> Please contact us
Note

Human serum or plasma often contains anti-HSV-1 antibodies making VirD inappropriate for clinical studies.

Sample Shipping and Sample Return

We will include shipping details in your quote – you must cover the cost of shipping samples to CDI Labs. Typically, after you receive your report, CDI Labs keeps the remaining samples for two months and then disposes of them. When shipping to us, please let us know if you want the remaining samples returned after the study is complete. Return shipping will be charged.