Creative Biolabs has successfully developed a full range of elegant antibody functional characterization assays to explore and validate the mechanism of action of antibody-based therapeutics. Particularly, we proudly bring out our well-recognized service of CDC detection assay.

Complement-dependent cytotoxicity, or CDC, is a well-known mechanism through which antibodies lyse the unwanted target by activating a cascade of complement-related reactions. Similar to ADCC, antibodies, usually IgG1 and IgG3, elicits CDC killing effects via binding its Fc region to serum complement components, particularly C1q. Following complicated enzyme activations and cleavage events, involving over 20 highly regulated elements, it will eventually lead to the formation of membrane attacking complex (MAC), thereby target cell destruction. Classical CDC manifests as one vital MOA of most therapeutic antibodies; meanwhile, it can collaborate with ADCC as an enhancer. Hence, detection, analysis, and quantification of CDC activity constitute an essential part of preclinical assessment and characterization of investigated antibodies.

In Creative Biolabs, our professional experts with profound experience have built an integrated CDC assay platform. Much alike with ADCC assay, we provide multiple choices for customers to select a most suitable format to meet each of the specific demands.

Mechanism of Complement-dependent Cytotoxicity Fig 1. Mechanism of Complement-dependent Cytotoxicity. (Boross P, Leusen JH. 2012)

Target Cells

We can deal with frozen cells from customers upon request. In case you haven't got your ideal target cell line, you can rely on our extensive expertise to find the most appropriate one. There are over a hundred well-qualified tumor cell lines available in Creative Biolabs. Our scientists also have vast experience in designing a variety of disease models.

Complement Sources

We offer high-quality complement system in human serum, which implements strict QC standards under regulatory guidelines. We can also provide serum from rodent/non-rodent species as an alternative.

Assay Endpoints

Multiple endpoints and readout formats are available in our laboratory. Typical endpoint by target cell lysis can be employed, e.g. LDH/ATP/GAPDH release, radioactivity measurement, as well as FACS screening. We can also design alternative assay readouts upon custom request.

Case Study
  • Objective: the study aims to characterize samples via CDC dose-response assay.
  • Assay Format:
    Target cells: Raji;
    Effector molecule: 5% NHSC;
    Dilution methods: starting concentration at 10ug/mL, dilution factor at 5-fold;

CDC concentration response of sample-X.Fig.2 CDC concentration response of sample-X. (Creative Biolabs)

Table.1 Best-fit values summary for the CDC dose-response study of sample-X. (Creative Biolabs)

log(agonist) vs. response -- Variable slope (four parameters)
Best-fit values
Bottom Top LogEC50 HillSlope EC50 Span
-1.751 73.07 -0.9951 1.28 0.1011 74.82

  • Summary:

In this study, results of the CDC dose-response study showed that the EC50 value of sample-X was 0.1011 μg/mL.

CDC response is a sophisticated process invoking a wide spectrum of cellular elements. Many parameters need to be considered to generate a highly relevant CDC assay. Creative Biolabs is willing to work with you step by step to tailor personalized experimental plans, including concentrations of target cells, determination of appropriate antibody/target ratio, sufficient antibody concentration range to yield a compelling dose-response curve. Aided by our unparalleled technique platform, Creative Biolabs is able to provide most robust, consistent outcomes with the assurance of the efficacy and potency of your antibody candidates. We also offer special Fc receptor binding assessment service as a further supplement of our immune effector function assays. For more detailed information, please feel free to contact us or directly sent us an inquiry.

Reference

  1. Boross P, Leusen JH. (2012) “Boosting antibody therapy with complement.” Blood. 119(25):5945-7.

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