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Related Concept Videos

Controlled-Current Coulometry: Coulometric Titration01:18

Controlled-Current Coulometry: Coulometric Titration

650
Coulometric titrations are a form of titrimetric analysis where the reagent is generated electrically, and its amount is evaluated based on current and generating time. The electron serves as the standard reagent. The procedure is similar to conventional titrations, such as endpoint detection.
The fundamental requirements for coulometric titrations are (1) 100% efficiency in the reagent-generating electrode reaction and (2) a stoichiometric and preferably rapid reaction between the generated...
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Titrimetric Methods: Types and Commonly Used Strategies01:08

Titrimetric Methods: Types and Commonly Used Strategies

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In chemistry, titrimetric methods are broadly classified into three types: volumetric, gravimetric, and coulometric. Volumetric titrations involve measuring the volume of a titrant of known concentration that is required to react completely with an analyte. In gravimetric titrations, the standard solution reacts with the analyte to form an insoluble precipitate, which is filtered, dried, and weighed. In coulometric titrations, current is applied to an electrochemical reaction until the reaction...
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EDTA: Direct, Back-, and Displacement Titration01:30

EDTA: Direct, Back-, and Displacement Titration

5.9K
The EDTA titration types for metal ion analysis include direct titration, back-titration, and replacement titration.
Direct titration involves buffering the metal ion solution to the desired pH and directly titrating with standard EDTA until the endpoint. The optimum pH ensures a large conditional formation constant of metal−EDTA and visibility of the free indicator color in the solution. In addition, auxiliary complexing reagents are used to prevent the precipitation of metal hydroxides...
5.9K
EDTA: Indirect and Alkalimetric Titration01:23

EDTA: Indirect and Alkalimetric Titration

2.0K
Unlike direct titration, back-titration, and displacement titration, indirect titration is an EDTA titration method for quantifying anions. In the indirect titration method, anions are precipitated as their insoluble salts with excess metal ions. The filtrate containing the excess metal ions is directly titrated with standard EDTA until the endpoint is achieved. Another approach involves extracting the metal ion and back-titrating with standard EDTA to obtain the endpoint. In this way, the...
2.0K
EDTA: Auxiliary Complexing Reagents01:26

EDTA: Auxiliary Complexing Reagents

1.5K
EDTA titrations are usually carried out in highly basic conditions, where the fully deprotonated form of EDTA, Y4−, actively complexes with the free metal ions in the solution. Several metal ions precipitate as hydrous oxide (hydroxides, oxides, or oxyhydroxides) under these conditions, lowering the concentration of free metal ions in the solution. For this reason, auxiliary complexing agents or ligands such as ammonia, tartrate, citrate, or triethanolamine are used in EDTA titrations to...
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A Semi-automated Approach to Preparing Antibody Cocktails for Immunophenotypic Analysis of Human Peripheral Blood
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Development of a Platform to Enable Fully Automated Cross-Titration Experiments.

Jason Cassaday1, Michael Finley1, Brian Squadroni1

  • 11 Screening & Protein Science, Merck and Co., North Wales, PA, USA.

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|November 20, 2016
PubMed
Summary
This summary is machine-generated.

This study introduces an automated system for drug discovery, enhancing the speed of mechanism of action (MOA) assays. The novel process significantly increases throughput for compound cross-titration experiments, accelerating lead optimization.

Keywords:
G protein–coupled receptorHTSagonist shiftallosteryautomated biologyengineeringhigh-throughput screeningrobotics and instrumentation

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Area of Science:

  • * Pharmacology
  • * Drug Discovery
  • * Biochemistry

Background:

  • * High-throughput screening (HTS) and lead optimization require compound potency and mechanism of action (MOA) determination.
  • * Traditional MOA assays, like agonist shift, are data-intensive, limiting throughput.
  • * Cross-titration experiments are crucial for MOA but are time-consuming.

Purpose of the Study:

  • * To enhance the throughput of compound cross-titration experiments for MOA determination.
  • * To automate MOA assays within a screening triage workflow.
  • * To enable rapid assessment of numerous compounds for G protein-coupled receptor (GPCR) modulation.

Main Methods:

  • * Integration of Hewlett Packard's D300 digital dispenser with a robotics platform.
  • * Development of an automated process for on-the-fly cross-titration in 1536-well plates.
  • * Utilization of in-house software and hardware for compound management, data tracking, and biological assays (IP1 and Ca2+ detection).

Main Results:

  • * Successful automation of cross-titration for positive and negative allosteric modulators.
  • * Application to two distinct GPCR targets using two different assay detection formats.
  • * Nearly 100 compounds tested per target, demonstrating a significant increase in assay throughput.

Conclusions:

  • * The developed automated system substantially enhances the efficiency of MOA assays.
  • * This platform accelerates the process of identifying and optimizing drug candidates.
  • * The technology is applicable to various GPCR targets and assay formats, facilitating drug discovery efforts.