Immune Response Assessment
When a pathogen infects the body, it initiates a cascade of complex immune reactions involving innate and adaptive arms of the immune system. Understanding these immune responses to infection provides greater insight as to why some individuals are more vulnerable to infection and related complications than others.
As COVID-19 has shown us, we need to gain deeper knowledge into the complex interplay between different elements of the immune system for rapid development of novel vaccine and therapeutic approaches.
Sartorius’ advanced cell and protein analysis platforms accelerate discovery and development of new vaccines against viral infections by providing comprehensive, more relevant data on key immune responses such as:
- Neutralizing antibodies
- T-Cell activation, memory and exhaustion
- Cytokine profiling
- Immune cell killing
- NETosis
- Vaccine titer and potency
Immune Response Categories
Neutralizing Antibodies
When a pathogen infects the body, it initiates a cascade of complex immune reactions involving innate and adaptive arms of the immune system. Understanding these immune responses to infection provides greater insight as to why some individuals are more vulnerable to infection and related complications than others.
Identification and Analysis of Neutralizing Antibodies with iQue Advanced Flow Cytometry Platform
Identifies and measures the affinity and kinetics of neutralizing antibodies to viral proteins.
The iQue® SARS-CoV-2 (IgG, IgM and IgA) Kit is a bead-based immunoassay designed for simultaneous detection of human IgG, IgM and IgA antibodies specific for SARS-CoV-2 Spike protein RBD produced by the immune system in response to viral infection or vaccination.
Figure 1: iQue® SARS-CoV-2 (IgG, IgM and IgA) Kit workflow. Easy to follow steps and low sample volumes needed (requires only a single 10 µL of plasma / serum diluted at least 100-fold).
Octet® Bio-Layer Interferometry Assays in Neutralization Antibodies Development Workflow 1
Epitope binning assays help identify antibodies that block the same epitope on a target antigen and are crucial when it comes to identifying or engineering mAbs with favorable kinetics and affinity profiles. This assay format can also be used to determine neutralization antibody competition with virus receptor binding to host receptors.
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Octet® Bio-Layer Interferometry Assays in Neutralization Antibodies Development Workflow 2
Real-time Kinetics (association and dissociation) and affinity data allows to confidently pick target binding lead candidates fast. Binding activity measurements can also be used to determine protein stability and activity, especially useful in vaccine antigen selection.
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Sartorius Immune Response Solutions Offer:
Live-cell analysis was used to quantitate real-time proliferation of Zika virus-infected and dengue virus-infected Vero cells treated with either Bardoxolne methyl, or Zika virus-neutralizing antiserum.
Combined phenotype and function assays including iQue® Qbeads® Kits capture specific proteins on distinct beads to facilitate multiplexed quantitation of a range of proteins, cytokines, enzymes and growth factors. The iQue® platform was used in conjunction with a highly sensitive bead-based fluorescent immunoassay from Sartorius for measuring SARS-CoV-2 specific antibody levels and isotypes in COVID-19 patient plasma and serum. Dogan M, Kozhaya L, Placek L, et al.
SARS-CoV-2 specific antibody and neutralization assays reveal the wide range of humoral immune response to virus.
Select the best vaccine and therapeutic candidates early in the development process based on binding affinity and kinetic assessments.
Octet® BLI instrument was used for receptor affinity characterization after BNT 162b2 RNA immunization in the study of B. Vogel et al, A prefusion SARS-CoV-2 spike RNA vaccine is highly immunogenic and prevents lung infection in non-human.
Product Highlights for Immune Response
iQue® SARS-CoV-2 (IgG, IgM and IgA) Kit
The iQue® SARS-CoV-2 (IgG, IgM and IgA) Kit contains ready-to-use reagents for simultaneous detection of human IgG, IgM and IgA antibodies specific for the receptor binding domain (RBD) of SARS-CoV-2 Spike protein in a single well using the iQue® VBR platform.
- Real-time binding kinetic and affinity measurements
- Measure interactions from purified and crude samples
- Fast data acquisition — simultaneously data from up to 96 biosensors
Incucyte® Live-Cell Analysis with Immune Cell Killing Assays
Modelling the dynamic interactions of immune- and virus-infected cells in vitro in real time. Gain new insight into immune cells, antibody function and killing using non-invasive, non-disruptive analysis platform of cell health, phenotype, subsets and function.
Octet BLI Assays in Neutralization Antibodies Development Workflows
Epitope binning assays help identify antibodies that block the same epitope on a target antigen and are crucial for identifying or engineering mAbs with favorable kinetics and affinity profiles.
Immune Response FAQs
The iQue automated, high-throughput, advanced flow cytometry platform allows scientists to rapidly multiplex protein analysis, immunophenotyping and perform functional assessments. It offers ready-to-use kits including the iQue® T Cell Phenotyping kit, Human Qbeads® Inflammation Panel for cytokine storm assessment, and kits for T cell activation, memory, killing, and exhaustion along with cytokine profiling for greater in-depth characterization of the host immune response in each sample.
Octet® BLI systems offer a rapid and effective method for protein quantitation due to their enhanced throughput, decreased sample preparation requirements, and low cost of operation. Samples run on Octet® systems are also recoverable, so that researchers may save and reuse precious sample for other experiments. In addition, Octet® assays are not affected by absorption interferences in colored samples or by light scattering with turbid samples, enabling measurement of analyte concentration in crude matrices such as cell culture supernatant, cell lysate and serum.
The iQue® advanced flow cytometry as well as the Octet® BLI systems have been widely used for measurement and characterization of humoral immune responses i.e. screening for antibody binding to SARS-CoV-2 proteins of COVID-19 patients.
Dogan, M., Kozhaya, L., Placek, L., Gunter, C., Yigit, M., Hardy, R., Plassmeyer, M., Coatney, P., Lillard, K., Bukhari, Z., Kleinberg, M., Hayes, C.,- Arditi, M., Klapper, E., Merin, N., Liang, B.T., Gupta, R., Alpan, O., & Unutmaz, D. (2021, Jan 29). SARS-CoV-2 specific antibody and neutralization assays reveal the wide range of the humoral immune response to virus. Commun Biol, 4(1), 129. https://doi.org/10.1038/s42003-021-01649-6 PMID: 33514825; PMCID: PMC7846565.
Zhang, G., Pomplun, S., Loftis, A.R., Loas, A., & Pentelute, B.L. (2020). The first-in-class peptide binder to the SARS-CoV-2 spike protein. bioRxiv, https://doi.org/10.1101/2020.03.19.999318
Wu, Y., Li, C., Xia, S., Tian, X., Kong, Y., Wang, Z., Gu, C., Zhang, R., Tu, C., Xie, Y., Yang, Z., Lu, L., Jiang, S., & Ying, T. (2020). Identification of Human Single-Domain Antibodies against SARS-CoV-2. Cell Host & Microbe, 27, 891–898. https://doi.org/10.1016/j.chom.2020.04.023
Live-cell based assays like virus-induced cell death, cell viability and cell-to-cell fusions assays as continuous real-time, live cell assays provide deeper insight into the cellular and functional interactions between immune cells, antibody responses and pathogens. Using the Incucyte® S3 Live-Cell Analysis System, researchers monitored cell death and cell-to-cell fusion when using isolated anti-SARS-CoV-2 antibodies from severe COVID-19 convalescent donors. Mor., M., Werbner, M., Alter, J., Safra, M., Chomsky, E., Lee J.C., et al. (2021). Multi-clonal SARS-CoV-2 neutralization by antibodies isolated from severe COVID-19 convalescent donors. PLoS Pathog, 17(2), e1009165.
Yes, you can combine the Incucyte® Annexin V Red Reagent with the Incucyte Cytotox Green Reagent for multiplexed measurements - readily discriminate between apoptosis and NETosis.
On-Demand Webinar: Evolution of Flow Cytometry in Infectious Diseases Research
Hear how our panelists study the various infection stages and the directions of their studies of infectious diseases.
