Antibody-dependent cellular cytotoxicity (ADCC) is an important immune mechanism for antibodies. For the past years, ADCC-enhanced therapeutic antibodies have been shown to exhibit unprecedented anti-cancer effects in vivo. During the research, characterizing ADCC activity and determining its impact on clinical efficacy is critical. With the leverage of our knowledge and rich experience in antibody engineering and glycosylation control, Creative Biolabs is committed to developing next-generation ADCC-enhanced therapeutic antibodies for our customers.
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FcγR Binding Assays
Octet-Based Kinetic & Affinity Characterization
Our ADCC assays can use human Peripheral blood mononuclear cell (PBMC) or NK cells as effector cells and the goal is to determine the antibody activity against specified target cell lines.
We have knowledge working with extensive cell banks (more than 100 qualified tumor cell lines). If you have trouble finding the right cell line to use, our technical support can help you find the most ideal cell line for your ADCC analysis.
PBMCs and NK cells are examples of classical immune cells involved in the ADCC effect, which both interact with the Fc portion of IgG via the Fc receptor (primarily FcγRIIIa).
Fig.1 Real-time monitoring of target cells at different time points.
Fig.2 In vitro ADCC assay.
Creative Biolabs offers a range of in-depth glycan analysis and Fc receptor binding assays to complement on your ADCC functional analysis to thoroughly characterize ADCC⁺ products.
We also provide ADCP assay service, please visit: ADCP Assay Services for more details.
Afucosylation of IgG N-glycans can influence receptor antibodies binding affinity, and thereby, acting on ADCC activity. A thorough characterization to determine the degree of antibody defucosylation is very important for ADCC⁺ therapeutics. We offer analytical methods for N-glycans in its different formats, from intact proteins, glycopeptides, released glycans, to individual monosaccharide components. Analysis characterization is done via high-resolution precision Hydrophilic interaction ultra-performance liquid chromatography (HILIC–UPLC).
Fig.3 Depending on the exact objective, our glycosylation analysis can be performed at different levels.
Creative Biolabs is able to perform FcγR binding assays on defucosylated Fc fusion proteins, using our Biacore's surface plasmon resonance (SPR) and Octet Biolayer Interferometry (BLI) technology platforms. The simplicity and utility of these two platforms for the determination of binding profiles of therapeutic ADCC⁺ antibodies and FcγR are aslo demonstrated during applciation.
With the help of experienced scientists, our FcγR binding assays are performed based on the following techniques:
Fig.4 Fc receptor binding assays on the Octet system.
Antibody aggregates are considered as impurities or molecular variants and may be caused by the effects of physical factors such as light, temperature, pH, water, shear force, or reaction with excipients over time in the formulation, etc. Common aggregates include charge variants, dimers, oligomers, or forced degradation variants of the desired antibody product.
|Antibody Purity Analysis||SDS-PAGE|
|Antibody Polymer Analysis||Size Exclusion Chromatography (SEC)|
|Antibody Fragment Analysis||Reversed-phase Chromatography (RP)|
|Antibody Isoform Analysis||Capillary isoelectric focusing|
Fig.5 Characterization of low molecular weight impurities associated with products in therapeutic monoclonal antibodies by hydrophilic interaction chromatography-mass spectrometry.
Biochemical and biophysical characterization of therapeutic antibodies to fully understand their structural and functional properties is extremely important to your research. Assessing Program Quality Assessment (PQA) is the foundation for product control strategies (specifications, comparability and stability strategies). Here at Creative Biolabs, we provide biochemical and biophysical characterization of APIs and pharmaceutical products during antibody drug development.
|Intact Mass Confirmation||Native, deglycosylated, and reduced protein electrospray ionization-mass spectrometry (ESI-MS)|
|Amino Acid Sequence Confirmation||MS/MS sequencing of peptide maps|
|N- and C-terminal Variants||MS/MS sequencing of peptide map|
|N- and C-terminal Sequencing||De novo antibody sequencing|
|Disulfide Structure||Non-reduced and reduced peptide map with MS Ellman's assay (or similar)|
|Secondary structure||Fourier transform infrared (FTIR) spectroscopy, circular dichroism|
|Tertiary structure||Near-UV circular dichroism (near-UV CD) spectroscopy, differential scanning calorimetry, X-ray crystallography|
|Thermal stability||Differential scanning calorimetry (DSC)|
Analysis of post-translational chemical modifications (including the categories and sites of modification) is particularly important for cell biology as well as clinical transformation. At Creative Biolabs, we offer various off-the-shelf chemical modification/ characterization services to assist your scientific research.
Creative Biolabs offers Octet System-based analysis services for antibody binding kinetic characterization. Octet complete kinetic analysis service allows the recording and identification of antibody drugs for their batch consistency and drug stability.
In addition to identifying binding partners to target molecules, Creative Biolabs also provides quantitative information regarding following properties:
Creative Biolabs is dedicated to developing the right bioassay to help you further your ADCC⁺ antibodies research. For more details about our Therapeutic ADCC⁺ Biobetter Antibody Analysis service, please do not hesitate to contact us.
Creative Biolabs provides luciferase-based ADCC assay. This Jurkat cell based assay is pioneered by Creative Biolabs, and the methodology is very well accepted by the field. See attached ADCC Reporter Assay Protocol for further details.
All products and services are for Research Use Only. Do Not use in humans.
ADCC Assay WT vs AfucoTM Mabs Visit
Antibody Fc Engineering: Towards Better Therapeutics Visit
Creative Biolabs has established a team of customer support scientists ready to discuss ADCC/CDC optimization strategies, antibody production, bioinformatics analysis and other molecular biology/biotechnology issues.