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Some receptors remain unoccupied even when an agonist produces a maximal response. Such empty ones are called spare receptors. In presence of spare receptors the maximum effect of an agonist drug is achieved with fewer than 100% of the receptors being occupied. To determine the presence of spare receptors, scientists often compare the concentration of the drug needed to produce 50% of the maximum effect (EC50) with the concentration of the drug needed to occupy 50% of the receptors (Kd). If the...
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In signal processing, the analysis of continuous-time signals, denoted as x(t), often involves sampling techniques to convert these signals into discrete-time signals. This process is essential for digital representation and manipulation. A critical component in sampling is the train of impulses, characterized by the sampling interval and the sampling frequency. The relationship between these parameters and the original signal's properties dictates the success of the sampling process.
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In signal processing, bandpass sampling is an effective technique for sampling signals that have most of their energy concentrated within a narrow frequency band. This type of signal is known as a bandpass signal. The key principle of bandpass sampling involves sampling the signal at a rate that is greater than twice the signal's bandwidth to prevent aliasing.
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Sampling Plans01:23

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Sampling is a crucial step in analytical chemistry, allowing researchers to collect representative data from a large population. Common sampling methods include random, judgmental, systematic, stratified, and cluster sampling.
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A Sample-Sparing Multiplexed ADCP Assay.

Audrey L Butler1, Jonathan K Fallon1, Galit Alter1

  • 1The Ragon Institute of MGH, MIT, and Harvard, Cambridge, MA, United States.

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|August 29, 2019
PubMed
Summary
This summary is machine-generated.

This study introduces a novel multiplexed assay for evaluating antibody functions against multiple antigens simultaneously. This bead-based method enhances efficiency and accuracy in analyzing antibody effector profiles, crucial for vaccine development.

Keywords:
antibody-mediated effector profilesdependent effector functionmonocytemultiplexphagocytosis

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Area of Science:

  • Immunology
  • Vaccinology
  • Assay Development

Background:

  • Antibodies are key correlates of vaccine protection, primarily through neutralization.
  • Antibody-mediated engagement of the innate immune system (phagocytosis, complement activation, cytotoxicity) is increasingly recognized as vital for protection.
  • Assessing these antibody effector functions is critical for vaccine trials, but requires high-throughput, sample-sparing methods.

Purpose of the Study:

  • To develop a robust, high-throughput, and sample-sparing assay for interrogating the functional capacity of innate immune-recruiting antibodies.
  • To enable simultaneous evaluation of antibody responses against multiple antigens from limited sample volumes.

Main Methods:

  • Adaptation of a single bead-based cellular assay to incorporate five different fluorescently colored beads.
  • Concurrent investigation of antibody responses against multiple antigens within a single assay well.
  • Validation of the multiplexed assay's sensitivity, specificity, and accuracy compared to single-antigen assays.

Main Results:

  • The developed multiplexed assay accurately assesses antibody functional activity against up to five antigen targets simultaneously.
  • The multiplexed approach demonstrated comparable sensitivity, specificity, and accuracy to single-antigen assays.
  • The assay robustly identified functional differences mediated by antibodies across various samples.

Conclusions:

  • Multiplexing bead-based cellular assays allows for efficient and accurate analysis of antibody-mediated effector profiles.
  • This approach is critical for profiling antibody functions across diverse pathogen antigens or variants, especially with limited sample volumes.
  • The developed method supports the identification of correlates of protection in large-scale vaccine trials.