At present, it is important to assess the potential immunogenicity of drugs during drug development to better guide clinical drug delivery while managing the clinical consequences of immunogenicity. Scientists tend to focus on developing the latest diagnostic and chemotherapeutic drugs, but these foreign substances can easily cause immunogenic mediated adverse effects in the subject. Therefore, the evaluation method of immunogenicity came into being. In this field, Creative Biolabs can provide a series of diagnostic services on immunogenicity, extensive information research, and quality technical support to ensure the smooth progress in your testing process.

Introduction of Real-time Kinetic Binding Assay

• Surface Plasmon Resonance (SPR)

The strength of immunogenicity is one of the determinants of biotechnology drug development, and immunogenicity testing is critical for evaluating drug safety. ELISA, western blot analysis and surface plasmon resonance SPR technology are all common methods for biopharmaceutical immunogenicity detection. Surface plasmon resonance (SPR) as one of the real-time kinetic binding assays, is to measure antigen-antibody interactions by "real-time" observation. When the sample flows through the antigen-fixed sensor chip, a continuous event signal is generated, and the signal changes as the refractive index changes. When an analyte binds to a ligand, the difference in mass causes a corresponding change in refractive index that is proportional to the amount (mass) of bound antibody in the sample being tested.

Research Application of Real-time Kinetic Binding Assay

TDIM was characterized by SPR to measure serum concentrations of infliximab, antibodies against tumor necrosis factor-alpha (anti-TNFa) and anti-infliximab antibodies. Some scientists have found that it can reduce mAbs interference when detecting low-affinity antibodies to therapeutic mAbs. SPR can directly detect and measure the value and content of serum antibodies in a short period, avoiding other cumbersome steps such as incubation, separation, and washing, while reducing the complexity of the data and the variability of factors. Besides, multiple drugs and their corresponding antibodies can be measured simultaneously. This method has been proven to have good sensitivity and has a higher profit than the ELISA kit.

Fig.1 General scheme of the SPR-based assay for simultaneous determination of IFX and ATI concentrations in serum. (Beeg, 2019)

Highlights

• Multiple methods for immunogenicity testing
• Extensive background knowledge
• Scientific test plan design

The real-time kinetic binding assay is suitable for the rapid analysis of data of various biotherapeutics and has broad application prospects. Creative Biolabs offers quality technical support of real-time kinetic binding assay and immunogenic detection. Please contact us as soon as possible and we are honored to serve you.

Published Data

• Real-Time Kinetic Analysis of Chemokine-Chemokine Receptor Interactions

Fig. 2 Nanodisc (ND) assembly and analysis. (Stefanie Alexandra Eberle, 2022)

The article discusses the development and application of a Real-Time Kinetic Binding Assay using a Scintillation Proximity Assay (SPA) to analyze interactions between chemokines and chemokine receptors, focusing on ACKR3 and CXCL12. This method is significant as it offers a detailed kinetic profile of these interactions, which is crucial for understanding the dynamics of chemokine receptor activity and their potential as drug targets. The results demonstrate that this SPA method can effectively measure real-time binding kinetics, providing both association and dissociation rates.

FAQ

1. What is a real-time kinetic binding assay?

   A real-time kinetic binding assay measures the interaction between a drug candidate and a target molecule over time, capturing both the association and dissociation rates to provide a detailed kinetic profile.

2. Why are kinetic parameters important in drug development?

   Kinetic parameters such as the association and dissociation rates provide critical insights into a drug's efficacy, potency, and duration of action, enabling more informed decisions in the drug development process.

3. Can real-time kinetic assays predict the clinical efficacy of a drug?

   While real-time kinetic assays provide valuable data on how a drug interacts with its target, predicting clinical efficacy also requires considering biological factors, such as drug metabolism and distribution, which are not captured in these assays.

4. How do real-time kinetic assays handle competitive binding scenarios?

   Real-time kinetic assays can be designed to assess competitive binding by introducing a competitor molecule. This setup helps in evaluating the potential of drug candidates to displace or be displaced by other ligands, which is crucial for understanding drug interactions.

5. What technologies are commonly used for real-time kinetic binding assays?

   Technologies such as surface plasmon resonance (SPR) and biolayer interferometry (BLI) are commonly used. These technologies detect changes in mass or optical properties at the sensor surface where the target is immobilized, allowing real-time monitoring of binding events.

6. What are the challenges of conducting real-time kinetic binding assays?

   Challenges include the need for high-quality, stable target proteins, accurate calibration of assay conditions, and interpreting complex data where multiple binding interactions may occur simultaneously.

Resources

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