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 Experiment Videos

A sensitive assay for maleimide groups

R Singh1

  • 1ImmunoGen, Inc., Cambridge, Massachusetts 02139.

Bioconjugate Chemistry
|July 1, 1994
PubMed
Summary
This summary is machine-generated.

A new spectrophotometric assay accurately quantifies maleimide groups in proteins. This sensitive method offers a 100-fold improvement over existing techniques for protein modification analysis.

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

Dietary intake, plasma levels of antioxidant vitamins, and oxidative stress in relation to coronary artery disease in elderly subjects.

The American journal of cardiology·1995
Same author

Role of endothelin-converting enzyme in the systemic hemodynamics and regional circulatory effects of proendothelin-1 (1-38) and diaspirin cross-linked hemoglobin in rats.

The Journal of laboratory and clinical medicine·1995
Same author

Randomized double-blind comparison of the analgesic efficacy of intramuscular ketorolac and oral indomethacin in the treatment of acute gouty arthritis.

Annals of emergency medicine·1995
Same author

Polymerase chain reaction-based genotyping of the constant segment of immunoglobulin kappa shows weak or no association with rheumatoid arthritis.

Arthritis and rheumatism·1995
Same author

Isolated oesophageal tuberculosis: a rare cause of oesophageal stricture.

The European journal of surgery = Acta chirurgica·1995
Same author

Distinct binding specificities and functions of higher eukaryotic polypyrimidine tract-binding proteins.

Science (New York, N.Y.)·1995
Same journal

Neutral Amphiphiles Boost Transfection Efficiency and Reduce Inflammation in Polymer Micelle-Mediated mRNA Delivery.

Bioconjugate chemistry·2026
Same journal

Surfactant-Mediated Buchwald-Hartwig Coupling of Aliphatic Amines for the Synthesis of DNA-Encoded Libraries.

Bioconjugate chemistry·2026
Same journal

Artificial Intelligence for Discovery in Life Sciences.

Bioconjugate chemistry·2026
Same journal

Iron Single Atom Nanozyme-Mediated GPX4 Inhibitor Delivery for Self-Enhanced Ferroptosis.

Bioconjugate chemistry·2026
Same journal

SpyCatcher-Engineered Ferritin Nanocages Enable Dual-Receptor Targeting for Enhanced Glioma Therapy.

Bioconjugate chemistry·2026
Same journal

One-Pot Synthesis of Functionalized Coumarin Fluorophores Enables Rapid Access to Live-Cell Bioorthogonal Labeling and Microenvironmental Sensing Agents.

Bioconjugate chemistry·2026
See all related articles

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Protein Chemistry

Background:

  • Maleimide groups are crucial for protein modification and bioconjugation.
  • Accurate quantification of maleimide is essential for characterizing modified proteins.
  • Existing assays, such as Ellman's reagent, may lack sufficient sensitivity.

Purpose of the Study:

  • To develop a highly sensitive spectrophotometric assay for quantifying maleimide groups in proteins.
  • To establish a reliable method for assessing protein modification with maleimides.
  • To provide a more sensitive alternative to current maleimide detection methods.

Main Methods:

  • Developed a spectrophotometric assay involving reaction of maleimide with excess cysteine.
  • Quantified remaining cysteine using an enzymatic assay based on papain activation.

Related Experiment Videos

  • Measured restored papain enzymatic activity with a chromogenic substrate.

    • Main Results:

    • The assay accurately determines maleimide quantity by calculating the difference between initial and remaining cysteine.
    • Achieved a limit of detection of approximately 0.1 nmol maleimide in a 1.2 mL final volume.
    • Demonstrated a 100-fold increase in sensitivity compared to assays using Ellman's reagent.

    Conclusions:

    • The developed assay is highly sensitive and accurate for quantifying maleimide groups in proteins.
    • This method provides a significant advancement in the analysis of maleimide-modified proteins.
    • The assay offers a valuable tool for researchers in protein chemistry and bioconjugation.