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Related Concept Videos

High-Performance Liquid Chromatography: Types of Detectors01:15

High-Performance Liquid Chromatography: Types of Detectors

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The role of the detectors in High-Performance Liquid Chromatography (HPLC) is to analyze the solutes as they exit from the chromatographic column. The detector recognizes the solute's property and generates corresponding electrical signals, which are converted into a readable graph of the detector's response versus elution time called a chromatogram at the computer. There are several types of HPLC detectors, each with its own advantages and limitations, depending on the analyte...
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High-Performance Liquid Chromatography: Introduction01:11

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High-performance liquid chromatography(HPLC), formerly referred to as High-pressure liquid chromatography, is a powerful technique used to separate, identify, and quantify components in complex mixtures. The term "high pressure" refers to using high pressure to push the liquid mobile phase through the tightly packed columns.
In HPLC, two phases play a critical role in the separation process:
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High-Performance Liquid Chromatography: Instrumentation00:57

High-Performance Liquid Chromatography: Instrumentation

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High-performance liquid chromatography, or HPLC, is an analytical technique that separates liquid samples under high pressures. An HPLC instrument consists of glass bottles for storing solvents called mobile phase reservoirs. HPLC-grade solvents are used to maintain high purity, and the dissolved gases are removed using a degasser, such as a vacuum pumping system or sparging with helium. The solvents are then pumped into the analytical column using a screw-driven syringe or reciprocating pumps.
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Mass Spectrometry: Complex Analysis01:21

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Mass spectrometry is an important technique for the identification of pure compounds. However, it has some limitations for the analysis of complex mixtures, often due to excessive fragmentation making the spectrum too complicated to decipher. Mass spectrometry can be combined with suitable separation methods in sequence, forming hyphenated methods, which are useful in the analysis of complex mixtures.
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
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Related Experiment Video

Updated: Jun 9, 2025

Author Spotlight: Detecting Low-Abundant Host Cell Proteins in Drug Products Using Enrichment Beads and Limited Digestion
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Analytical Tools for HCP Detection, Identification, and Quantitation.

Faiza Kanwal1, Michael Henry1, Paula Meleady2,3

  • 1National Institute for Cellular Biotechnology, Dublin City University, Dublin, Ireland.

Methods in Molecular Biology (Clifton, N.J.)
|October 26, 2024
PubMed
Summary

Host cell protein (HCP) analysis is vital for biopharmaceutical safety. Advanced liquid chromatography-mass spectrometry (LC-MS/MS) offers comprehensive HCP identification and quantification, improving process control and product safety.

Keywords:
Biopharmaceutical manufacturingData acquisition modesData analysis strategiesHost cell proteinsLC-MS/MSSample preparation

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An HS-MRM Assay for the Quantification of Host-cell Proteins in Protein Biopharmaceuticals by Liquid Chromatography Ion Mobility QTOF Mass Spectrometry
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An HS-MRM Assay for the Quantification of Host-cell Proteins in Protein Biopharmaceuticals by Liquid Chromatography Ion Mobility QTOF Mass Spectrometry

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Area of Science:

  • Biopharmaceutical Manufacturing
  • Analytical Chemistry
  • Proteomics

Background:

  • Host cell proteins (HCPs) are critical impurities in biopharmaceutical products.
  • Enzyme-linked immunosorbent assay (ELISA) is a traditional method for HCP detection.
  • Limitations exist in ELISA for comprehensive HCP profiling.

Purpose of the Study:

  • To explore advancements in liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) for HCP analysis.
  • To discuss challenges and innovations in HCP identification and quantitation.
  • To highlight the potential of LC-MS/MS in biopharmaceutical process control and safety.

Main Methods:

  • Review of LC-MS/MS methodologies for HCP analysis.
  • Discussion of sample preparation techniques.
  • Examination of data acquisition and analysis strategies.

Main Results:

  • LC-MS/MS provides comprehensive and quantitative data on individual HCPs.
  • Recent innovations address limitations in HCP identification and quantitation.
  • LC-MS/MS serves as an orthogonal tool to ELISA.

Conclusions:

  • LC-MS/MS methods are advancing HCP analysis in biopharmaceuticals.
  • Improved HCP profiling enhances process understanding and control.
  • LC-MS/MS application promises increased product safety in biopharmaceutical manufacturing.