Lumit® FcγR Binding Immunoassays
Measure Fcγ Receptor Binding
- No wash or transfer; simple add-mix-read protocol
- Results in less than 1 hour
- Scalable and high throughput; perform assays in 96-well or 384-well plates
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Catalog Number: W7030
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$ 730.00
Catalog Number: W7050
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Catalog Number: W7080
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Faster, Simpler Fc Receptor Binding Assays
The Lumit® FcγR Binding Immunoassays are novel homogeneous (no-wash) competition assays to measure the interaction between human Fc receptors and antibodies or Fc fusion proteins. Lumit® Immunoassays are based on NanoBiT® complementation to generate a luminescent signal. In the assays, a human IgG labeled with LgBiT (Tracer-LgBiT) is used as the tracer. A human FcγR labeled with SmBiT is used as the target. In the absence of an antibody analyte sample, Tracer-LgBiT binds to the FcγR-SmBiT target, resulting in maximum luminescence signal. In analyte samples, unlabeled IgG will compete with Tracer-LgBiT for binding to the FcγR target, resulting in a concentration-dependent decrease in luminescent signal. These easy-to-use biochemical assays can complement and provide orthogonal data to support results from cell-based functional bioassays.
Fcγ Receptor Function
Fc receptors for IgG (FcγRs) are expressed by a variety of cells of hematopoietic origin and contribute to protective functions of the immune system. These receptors are so named because they possess affinity for the fragment-crystallizable (Fc) region of antibody molecules. Upon binding antibodies attached to diseased cells or pathogens, these receptors mediate effector functions such as antibody-dependent cellular cytotoxicity (ADCC) and antibody-dependent cellular phagocytosis (ADCP). The various Fcγ receptors differ in their affinity for IgG subclasses, and these interactions are further modified by the IgG glycosylation pattern.
Note: The total number of assays per kit will vary based on final well volume.
For standard well volumes:
- 100 assay kits: applicable for 100 assays in 96-well plates or 400 assays in 384-well plates.
- 1,000 assay kits: applicable for 1,000 assays in 96-well plates or 4,000 assays in 384-well plates.
Learn more about the Lumit® FcγR Binding Immunoassays
Nath, N. et al. (2022) A homogeneous bioluminescent immunoassay for parallel characterization of binding between a panel of antibodies and a family of Fcγ receptors. Sci Rep 12, 12185.
Simple Protocol, Reproducible Results
The Lumit® FcγR Binding Immunoassays are solution-based homogeneous assays that require no washes or sample transfer steps.
How the Lumit® FcγR Binding Immunoassays Work
In the absence of an antibody analyte, Tracer-LgBiT binds to the FcγR-SmBiT target, resulting in maximum luminescence signal. In samples containing analyte, nonlabeled IgG will compete with Tracer-LgBiT for binding to the FcγR target, resulting in a concentration-dependent decrease in the luminescent signal.
Assay Performance Data
Differential binding of anti-hCD20 isotypes to the high-affinity FcγRI measured with the Lumit® FcγRI Binding Immunoassay. A representative binding curve for anti-hCD20 IgG1, IgG2, IgG3 and IgG4 interacting with FcγRI is shown as normalized luminescence. The maximal bioluminescent signal generated in the absence of analyte (0nM antibody) was defined as 100%, and the signal at each antibody concentration was expressed as a percentage of this maximum. Data were fit with a four-parameter logistic regression model using 1/y² weighting to determine IC₅₀ values.
FcγRIIIa (V158) binding of rituximab biosimilars measured with the Lumit® FcγRIIIa (V158) Binding Immunoassay. A representative binding curve for Control Anti-CD20 IgG1, ublituximab and obinutuzumab is plotted as normalized luminescence. The maximal bioluminescent signal observed in the absence of analyte (0nM antibody) was defined as 100%, and the signal at each antibody concentration was expressed as a percentage of this maximum. Data were fitted to a four-parameter logistic regression model with 1/y² weighting to calculate apparent IC₅₀ values.
Effect of H₂O₂-mediated oxidation on FcγRIIa (H131) binding of human IgG3 measured with the Lumit® FcγRIIa Binding Immunoassay. Human IgG3 was added to H₂O₂ solutions to yield final peroxide concentrations of 3%, 1%, 0.3% or 0% (control). Samples were exposed for 2 hours at RT followed by removal of residual H₂O₂. Serial dilutions of each treated antibody were assayed, and raw luminescence (RLU) was plotted against IgG3 concentration. Curves were fitted with a four-parameter logistic regression model employing 1/y² weighting to determine apparent IC₅₀ values.
Effect of Fc glycan removal on FcγRI binding measured with the Lumit® FcγRI Binding Immunoassay. A representative binding curve is shown for native human IgG and PNGase F–deglycosylated IgG. Deglycosylation was performed overnight at 37°C (4mg/ml⁻¹ IgG; 1ml PNGase F per 20mg of IgG). The maximal bioluminescent signal obtained in the absence of analyte (0nM IgG) was defined as 100%, and the signal at each IgG concentration was expressed as a percentage of this maximum. Data were fitted to a four-parameter logistic regression equation with 1/y² weighting to calculate apparent IC₅₀ values.
Lumit® FcγRIIb Binding Immunoassay. A representative standard curve of FcγR Control Antibody is plotted as normalized data. Normalized luminescence is calculated by setting the maximum bioluminescent signal (observed in the absence of an analyte) as 100% and expressing the signal in the presence of an analyte as a percentage of this maximum. Data were fitted to a four-parameter logistic regression equation with 1/y2 weighting.
Protocols
Complete Protocol
Specifications
Catalog Number:
What's in the box?
| Item | Part # | Size |
|---|---|---|
Lumit® Detection Substrate A |
VB301D | 1 × 75μl |
Tracer-LgBiT |
W111A | 1 × 60μl |
FcγRIIb-SmBiT |
W703A | 1 × 40μl |
FcγR Assay Buffer |
W709A | 1 × 25ml |
FcγR Control Antibody |
W710A | 1 × 120μl |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
U.S. Pat. Nos. 9,797,889, 9,797,890, 10,107,800 and 11,493,504; European Pat. No. 2970412; Japanese Pat. Nos. 7280842 and 7532562; and other patents and patents pending.
What's in the box?
| Item | Part # | Size |
|---|---|---|
Lumit® Detection Substrate A |
VB301D | 10 × 75μl |
Tracer-LgBiT |
W111A | 10 × 60μl |
FcγRIIb-SmBiT |
W703A | 10 × 40μl |
FcγR Assay Buffer |
W709A | 10 × 25ml |
FcγR Control Antibody |
W710A | 10 × 120μl |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.U.S. Pat. Nos. 9,797,889, 9,797,890, 10,107,800 and 11,493,504; European Pat. No. 2970412; Japanese Pat. Nos. 7280842 and 7532562; and other patents and patents pending.
What's in the box?
| Item | Part # | Size |
|---|---|---|
Lumit® Detection Substrate A |
VB301D | 1 × 75μl |
Tracer-LgBiT |
W111C | 1 × 120μl |
FcγRIIIa (F158)-SmBiT |
W704A | 1 × 30μl |
FcγR Assay Buffer |
W709A | 1 × 25ml |
FcγR Control Antibody |
W710A | 1 × 120μl |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
U.S. Pat. Nos. 9,797,889, 9,797,890, 10,107,800 and 11,493,504; European Pat. No. 2970412; Japanese Pat. Nos. 7280842 and 7532562; and other patents and patents pending.
What's in the box?
| Item | Part # | Size |
|---|---|---|
Lumit® Detection Substrate A |
VB301D | 10 × 75μl |
Tracer-LgBiT |
W111C | 10 × 120μl |
FcγRIIIa (F158)-SmBiT |
W704A | 10 × 30μl |
FcγR Assay Buffer |
W709A | 10 × 25ml |
FcγR Control Antibody |
W710A | 10 × 120μl |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.U.S. Pat. Nos. 9,797,889, 9,797,890, 10,107,800 and 11,493,504; European Pat. No. 2970412; Japanese Pat. Nos. 7280842 and 7532562; and other patents and patents pending.
What's in the box?
| Item | Part # | Size |
|---|---|---|
Lumit® Detection Substrate A |
VB301D | 1 × 75μl |
Tracer-LgBiT |
W111A | 1 × 60μl |
FcγRIIIa (V158)-SmBiT |
W705A | 1 × 20μl |
FcγR Assay Buffer |
W709A | 1 × 25ml |
FcγR Control Antibody |
W710A | 1 × 120μl |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
U.S. Pat. Nos. 9,797,889, 9,797,890, 10,107,800 and 11,493,504; European Pat. No. 2970412; Japanese Pat. Nos. 7280842 and 7532562; and other patents and patents pending.
What's in the box?
| Item | Part # | Size |
|---|---|---|
Lumit® Detection Substrate A |
VB301D | 10 × 75μl |
Tracer-LgBiT |
W111A | 10 × 60μl |
FcγRIIIa (V158)-SmBiT |
W705A | 10 × 20μl |
FcγR Assay Buffer |
W709A | 10 × 25ml |
FcγR Control Antibody |
W710A | 10 × 120μl |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.U.S. Pat. Nos. 9,797,889, 9,797,890, 10,107,800 and 11,493,504; European Pat. No. 2970412; Japanese Pat. Nos. 7280842 and 7532562; and other patents and patents pending.
What's in the box?
| Item | Part # | Size |
|---|---|---|
Lumit® Detection Substrate A |
VB301D | 1 × 75μl |
Tracer-LgBiT |
W111A | 1 × 60μl |
FcγRIIa (R131)-SmBiT |
W706A | 1 × 20μl |
FcγR Assay Buffer |
W709A | 1 × 25ml |
FcγR Control Antibody |
W710A | 1 × 120μl |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
U.S. Pat. Nos. 9,797,889, 9,797,890, 10,107,800 and 11,493,504; European Pat. No. 2970412; Japanese Pat. Nos. 7280842 and 7532562; and other patents and patents pending.
What's in the box?
| Item | Part # | Size |
|---|---|---|
Lumit® Detection Substrate A |
VB301D | 10 × 75μl |
Tracer-LgBiT |
W111A | 10 × 60μl |
FcγRIIa (R131)-SmBiT |
W706A | 10 × 20μl |
FcγR Assay Buffer |
W709A | 10 × 25ml |
FcγR Control Antibody |
W710A | 10 × 120μl |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.U.S. Pat. Nos. 9,797,889, 9,797,890, 10,107,800 and 11,493,504; European Pat. No. 2970412; Japanese Pat. Nos. 7280842 and 7532562; and other patents and patents pending.
What's in the box?
| Item | Part # | Size |
|---|---|---|
Lumit® Detection Substrate A |
VB301D | 1 × 75μl |
Tracer-LgBiT |
W111A | 1 × 60μl |
FcγRIIa (H131)-SmBiT |
W707A | 1 × 20μl |
FcγR Assay Buffer |
W709A | 1 × 25ml |
FcγR Control Antibody |
W710A | 1 × 120μl |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
U.S. Pat. Nos. 9,797,889, 9,797,890, 10,107,800 and 11,493,504; European Pat. No. 2970412; Japanese Pat. Nos. 7280842 and 7532562; and other patents and patents pending.
What's in the box?
| Item | Part # | Size |
|---|---|---|
Lumit® Detection Substrate A |
VB301D | 10 × 75μl |
Tracer-LgBiT |
W111A | 10 × 60μl |
FcγRIIa (H131)-SmBiT |
W707A | 10 × 20μl |
FcγR Assay Buffer |
W709A | 10 × 25ml |
FcγR Control Antibody |
W710A | 10 × 120μl |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.U.S. Pat. Nos. 9,797,889, 9,797,890, 10,107,800 and 11,493,504; European Pat. No. 2970412; Japanese Pat. Nos. 7280842 and 7532562; and other patents and patents pending.
What's in the box?
| Item | Part # | Size |
|---|---|---|
Lumit® Detection Substrate A |
VB301D | 1 × 75μl |
Tracer-LgBiT |
W111B | 1 × 10μl |
FcγRI-SmBiT |
W708A | 1 × 25μl |
FcγR Assay Buffer |
W709A | 1 × 25ml |
FcγR Control Antibody |
W710A | 1 × 120μl |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
U.S. Pat. Nos. 9,797,889, 9,797,890, 10,107,800 and 11,493,504; European Pat. No. 2970412; Japanese Pat. Nos. 7280842 and 7532562; and other patents and patents pending.
What's in the box?
| Item | Part # | Size |
|---|---|---|
Lumit® Detection Substrate A |
VB301D | 10 × 75μl |
Tracer-LgBiT |
W111B | 10 × 10μl |
FcγRI-SmBiT |
W708A | 10 × 25μl |
FcγR Assay Buffer |
W709A | 10 × 25ml |
FcγR Control Antibody |
W710A | 10 × 120μl |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.U.S. Pat. Nos. 9,797,889, 9,797,890, 10,107,800 and 11,493,504; European Pat. No. 2970412; Japanese Pat. Nos. 7280842 and 7532562; and other patents and patents pending.
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