Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

The life history of recessive deleterious alleles as seen through the eyes of a honey bee (Apis mellifera).

Molecular biology and evolution·2026
Same author

Immune effector cell-associated hematotoxicity following CAR-T cell therapies: insights from a large national database.

Japanese journal of clinical oncology·2026
Same author

Co-culturing hiPSC-cardiomyocytes and cardiac fibroblasts enhances engineered heart tissue structure and function.

Stem cells translational medicine·2026
Same author

Enzymatic Deacetylation as a Selective Strategy for <i>O</i>-GlcNAc Identification.

Analytical chemistry·2026
Same author

Characterizing the post-market safety profile of cemiplimab: a pharmacovigilance study of the FDA adverse events reporting system database.

Investigational new drugs·2026
Same author

CRS and ICANS as class-level adverse event for CD20 ×CD3 bispecific T-cell engagers: A Bayesian analysis of a nationwide database.

Leukemia research·2026

Related Experiment Video

Updated: Jun 24, 2025

Digital Microfluidics for Automated Proteomic Processing
10:55

Digital Microfluidics for Automated Proteomic Processing

Published on: November 6, 2009

12.5K

Development of automated proteomic workflows utilizing silicon-based coupling agents.

Connor Frey1, Maor Arad2, Kenneth Ku3

  • 1Department of Chemistry, University of the Fraser Valley, 33844 King Road, Abbotsford, BC V2S 7M8, Canada; Faculty of Medicine, University of British Columbia, 2194 Health Sciences Mall, Vancouver, BC V6T 1Z3, Canada.

Journal of Proteomics
|June 6, 2024
PubMed
Summary
This summary is machine-generated.

New silicone-based coupling agents, 4-triethoxysilylbutyraldehyde (TESB) and TESBA, enable automated enzyme immobilization for robust, rapid digestion in proteomics. This method streamlines sample preparation and reduces enzyme waste for cost-effective analysis.

Keywords:
Automated liquid handlerBottom-up proteomicsCapillary electrophoresisImmobilized enzymeLiquid-chromatography mass spectrometryPeptide mapping

More Related Videos

A Plasma Sample Preparation for Mass Spectrometry using an Automated Workstation
07:12

A Plasma Sample Preparation for Mass Spectrometry using an Automated Workstation

Published on: April 24, 2020

10.0K
Attaching Biological Probes to Silica Optical Biosensors Using Silane Coupling Agents
09:35

Attaching Biological Probes to Silica Optical Biosensors Using Silane Coupling Agents

Published on: May 1, 2012

13.0K

Related Experiment Videos

Last Updated: Jun 24, 2025

Digital Microfluidics for Automated Proteomic Processing
10:55

Digital Microfluidics for Automated Proteomic Processing

Published on: November 6, 2009

12.5K
A Plasma Sample Preparation for Mass Spectrometry using an Automated Workstation
07:12

A Plasma Sample Preparation for Mass Spectrometry using an Automated Workstation

Published on: April 24, 2020

10.0K
Attaching Biological Probes to Silica Optical Biosensors Using Silane Coupling Agents
09:35

Attaching Biological Probes to Silica Optical Biosensors Using Silane Coupling Agents

Published on: May 1, 2012

13.0K

Area of Science:

  • Biochemistry and Proteomics
  • Analytical Chemistry
  • Materials Science

Background:

  • Enzyme immobilization is crucial for efficient and reusable proteomic sample preparation.
  • Classical methods can be time-consuming and labor-intensive.
  • Development of novel coupling agents and automated workflows is needed for high-throughput proteomics.

Purpose of the Study:

  • To develop automated methods for enzyme immobilization using novel silicone-based coupling agents.
  • To evaluate the efficiency and robustness of immobilized enzyme particles (IEPs) and microreactors (IMERs).
  • To adapt these methods for streamlined, high-throughput bottom-up proteomics (BUP).

Main Methods:

  • Development and application of 4-triethoxysilylbutyraldehyde (TESB) and 4-triethoxysilylbutanoic acid (TESBA) as silicone-based enzyme coupling agents.
  • Automated fabrication of immobilized enzyme particles (IEPs) using an Agilent Bravo liquid handling system.
  • Adaptation of the IEP method for in-situ immobilized enzyme microreactor (IMER) fabrication within silica capillaries.

Main Results:

  • IEPs demonstrated robustness, rapid digestion, and high immobilization efficiency (51 ± 8%).
  • Automated IEPs using trypsin rivaled classical in-gel digestion; pepsin IEPs showed specific cleavage at leucine residues.
  • IMERs achieved efficient digestion of bovine serum albumin (BSA) with 33-40% sequence coverage in 15 min.

Conclusions:

  • TESB and TESBA offer effective silicone-based solutions for enzyme immobilization.
  • Automated IEP and IMER methods significantly enhance efficiency, reproducibility, and cost-effectiveness in proteomics.
  • These advancements pave the way for fully automated, mass spectrometry-integrated proteomics workflows.