Immunomodulatory Compound Screening

PRR Ligand Screening Service

TLR screening

InvivoGen offers screening services to detect compounds that activate or block the innate immune system:

- TLR Ligand Screening
- NOD1/2 Ligand Screening
- RIG-I/MDA-5 Ligand Screening
- Dectin-1 Ligand Screening
- Mincle Ligand Screening

Over the past several years InvivoGen has developed a large collection of cellular assays to detect compounds that activate or block the immune system through activation of PRRs with an emphasis on TLRs, NOD1/2 and RIG-I/MDA-5.

These sensitive cellular assays feature engineered HEK293 cells which utilize an NF-kB-inducible SEAP (secreted embryonic alkaline phosphatase) or an IRF (interferon regulatory factor)-inducible sLUC (secreted luciferase) reporter gene as the read-out. Ligand-induced activation of the NF-kB or IRF pathways is monitored using proprietary detection assays designed to provide rapid and reliable results.

Specifications

Short turnaround time - Screening turnaround: ONLY 3 weeks

Screening flexibility - Screening parameters can be selected and/or modified based on customer requirements.

Cost effective - A set-up charge applies for the first compound. Subsequent compounds are heavily discounted.

Reliable - Our screening service has been utilized consistently by leading Biotech and Pharmaceutical companies and academic institutes for many years.

Contents

Screening Services

Two choices of services are offered that can be performed sequentially or individually :

• Compound Profiling (level 1): Single dose testing on a set of PRRs.
Screening is performed at a single concentration, typically a 1/10 dilution of the original compound solution provided, or customer specified.

• Compound Dose Response (level 2): Dose response on one or several PRRs
Three concentrations of the compound(s), typically 1/10, 1/100 and 1/1000 dilutions of the original compound solution, are tested on the PRR(s) recognizing the compound(s) as determined in level 1 or specified by the customer.

A detailed report is prepared and provided to the customer electronically and in hard copy. All procedures are performed accordingly to strict guidelines. Confidentiality is guaranteed.

Description

TLR Ligand Screening

The TLR ligand screening service utilizes the HEK-Blue TLR cells.
These HEK293-derived cells are NF-kB-SEAP repor ter cells that stably express a human or mouse TLR gene. The TLR genes expressed are TLR2, 3, 4, 5, 7, 8, and 9.
Compound profiling: The compound is tested on all human and/or mouse TLRs to determine which TLR recognizes this compound. TLR ligands are typically recognized by a single TLR or potentially two (TLR7 and TLR8). Recognition by TLR4 usually reflects the presence of endotoxins in the sample.
Agonist and antagonist assays can be performed.

NOD1/2 Ligand Screening

The NOD1/2 ligand screening service utilizes the HEK-Blue NOD cells.
These HEK-Blue cells stably express an NF-κB-inducible SEAP reporter gene and the human or mouse NOD1 or NOD2 gene.
The compound can be tested on human and/or mouse NOD1 and NOD2.

RIG-I/MDA-5 Ligand Screening

The RIG-I/MDA-5 ligand screening service utilizes HEK293 cell lines that exploit the IRF pathway and a secreted luciferase reporter assay. They overexpress the human RIG-I or MDA-5 gene and are highly sensitive to 5’ppp-dsRNA and transfected poly(I:C), respectively, unlike their parental cell line that expresses low levels of both RIG-I and MDA-5.
The compound can be tested on human RIG-I and/or MDA-5.

Dectin-1 Ligand Screening

The Dectin-1 ligand screening service utilizes the HEK-Blue™ Dectin-1 cells.
These cell lines express different isoforms of the Dectin-1 gene. They also express genes of the Dectin-1-NF-κB signalling pathway, in addition to an NF-κB-inducible SEAP reporter gene.
The coumpound can be tested on human Dectin-1a, human Dectin-1b and/or mouse Dectin-1a.
Agonist and antagonists assays can be performed.

Mincle Ligand Screening

The Mincle ligand screening service utilizes the HEK-Blue™ hMincle cell line.
This cell co-expresses the human Mincle gene, genes of the Mincle-NF-κB signaling pathway and an NF-κB-inducible SEAP reporter gene.Agonist and antagonists assays can be performed, although as of today no ligand with inhibitory activity on Mincle has been identified.

 

Details

Simplified representation of the TLR, NOD and RLR pathways

Simplified representation of the TLR, NOD and RLR pathways

Citations

Recent articles using Immunomodulatory Compound Screening

ORDERING

Compound Profiling

Description Single dose testing on a set of PRRs
Cat. Code tlrl-test1
Unit Size 1 test
Price For price or distributor address,
please select your country

    Compound Dose Response

    Description Dose response on one or several PRRs
    Cat. Code tlrl-test2
    Unit Size 1 test
    Price For price or distributor address,
    please select your country

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      Note that the sequence of available ORFs provided by InvivoGen can differ from a given reference Genbank record due to genetic variations and/or alternative splicing. Customers should verify that the version of a gene sold by InvivoGen is suitable for the customer needs.
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