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.
Target Cell
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.
Effector Cells
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).
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Primary ADCC Assay—Creative Biolabs collects fresh PBMC or purified NK cells from healthy
genotypes from healthy or diseased donors using standard procedures.
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ADCC Report Analysis—ADCC analysis based on primary effector cells can be both time consuming and
expensive. Also, using PBMC is reported to have intra- and inter-assay variability, which may lead
to inconsistent results. Therefore, Creative Biolabs uses engineered effector cells to react with
the FcγRIIIa receptor and expresses FcγRIIIa and luciferase, which allows for optimal analytical
performance by ADCC reporting bioassays.
In addition, Creative Biolabs offers a range of stable cell lines genetically engineered to provide your ADCC/ADCP functional validation assays. We transduced a Luciferase reporter driven by NFAT signaling in Jurkat cells and simultaneously expressed FcγR on the cell membrane surface. When the cell is co-cultured with the target cell and the associated antibody, the antibody binds to both the target cell antigen and the cell surface Fcγ receptor, resulting in receptor aggregation, intracellular signaling, and NFAT-mediated Luciferase reporter gene expression and luminescence.
Analytical Endpoint
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Target cell lysis in vivo and in vitro: LDH (lactate dehydrogenase) assay / ATP release /
FACS screening
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Effector cell activation: luminescent NFAT reporter gene
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Real-time monitoring: our platform actively analyzes target cell proliferation and death rate in
the presence of effector cells and test antibodies, rather than single-end measurement. This allows
for a quick, simple but sensitive detection of subtle ADCC activity.
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.
Glycan Analysis
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).
Fc Receptor Binding (FcγR) Assays
Creative Biolabs is able to perform FcγR binding assays on defucosylated Fc fusion proteins, using our
surface plasmon resonance and Biolayer Interferometry technology platforms.
The simplicity and utility of these two platforms for the determination of binding profiles of
therapeutic ADCC⁺ antibodies and FcγR are also demonstrated during application.
With the help of experienced scientists, our FcγR binding assays are performed based on the following
techniques:
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Conventional ELISA
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Flow cytometry
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Surface Plasmon Resonance (SPR)/ Octet Biolayer Interferometry (BLI) (recommended): give real-time
unlabeled full kinetic analysis, and able to report affinity constants (KD) associated with detailed
or "on" rates (Ka), dissociation rates (Kd) and/or "closed" rates
Aggregation Characterization
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.
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Services
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Techniques
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Antibody Purity Analysis
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SDS-PAGE
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Antibody Polymer Analysis
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Size Exclusion Chromatography (SEC)
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Antibody Fragment Analysis
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Reversed-phase Chromatography (RP)
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Antibody Isoform Analysis
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Capillary isoelectric focusing
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Physicochemical Characterization
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 active pharmaceutical ingredient (APIs) and pharmaceutical products during antibody drug development.
Primary Structure Characterization
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Services
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Techniques
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Intact Mass Confirmation
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Native, deglycosylated, and reduced protein electrospray ionization-mass spectrometry
(ESI-MS)
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Amino Acid Sequence Confirmation
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MS/MS sequencing of peptide maps
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N- and C-terminal Variants
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MS/MS sequencing of peptide map
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N- and C-terminal Sequencing
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De novo antibody sequencing
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Disulfide Structure
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Non-reduced and reduced peptide map with MS Ellman's assay (or similar)
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Biophysical Characterization
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Services
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Techniques
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Secondary structure
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Fourier transform infrared (FTIR) spectroscopy, circular dichroism
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Tertiary structure
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Near-UV circular dichroism (near-UV CD) spectroscopy, differential scanning calorimetry,
X-ray crystallography
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Thermal stability
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Differential scanning calorimetry (DSC)
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Chemical Modification Characterization
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.
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Glycosylation Analysis
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Phosphorylation Analysis
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Deamidation and Oxidation Analysis
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Disulfide Bridges & Free Sulfhydryl Groups
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Antibody C-terminal Lysine Variants Determination
Octet-Based Kinetic & Affinity Characterization
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.
Available Octet Systems
In addition to identifying binding partners to target molecules, Creative Biolabs also provides
quantitative information regarding following properties:
Specificity
Concentration
Kinetics
Affinity
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.
