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

Simultaneous multiple analyte detection using fluorescent peptides and capillary isoelectric focusing

K A Cruickshank1, J Olvera, U R Muller

  • 1Vysis Inc., Downers Grove, IL 60515, USA.

Journal of Chromatography. A
|October 9, 1998
PubMed
Summary

Researchers developed fluorescent synthetic peptides for analyte detection using capillary isoelectric focusing-laser-induced fluorescence (cIEF-LIF). This method achieves picomolar detection limits and simultaneous analysis of multiple analytes.

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

Specific Immunotherapy in Hymenoptera Venom Allergy and Concomitant Malignancy: A Retrospective Follow-up Focusing on Effectiveness and Safety.

Journal of investigational allergology & clinical immunology·2017
Same author

Uterine contractions are known side effects of venom immunotherapy.

Journal of investigational allergology & clinical immunology·2011
Same author

Influence of total and specific IgE, serum tryptase, and age on severity of allergic reactions to Hymenoptera stings.

Allergy·2010
Same author

Elevated baseline serum tryptase, mastocytosis and anaphylaxis.

Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology·2009
Same author

Hymenoptera venom allergy: analysis of double positivity to honey bee and Vespula venom by estimation of IgE antibodies to species-specific major allergens Api m1 and Ves v5.

Allergy·2009
Same author

Intracranial germ cell tumors: association with Klinefelter syndrome and sex chromosome aneuploidies.

Cytogenetic and genome research·2008

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Biotechnology

Background:

  • Analyte detection methods are crucial in various scientific fields.
  • Existing methods like capillary electrophoresis-laser-induced fluorescence (CE-LIF) have limitations in detection sensitivity.
  • Novel approaches are needed for more precise and sensitive bioanalyte quantification.

Purpose of the Study:

  • To investigate a new concept for analyte-specific detection based on the isoelectric point of a detection moiety.
  • To develop and validate a method for synthesizing fluorescent synthetic peptides for bioanalyte conjugation and detection.
  • To establish a sensitive and reproducible detection system using capillary isoelectric focusing with laser-induced fluorescence (cIEF-LIF).

Main Methods:

  • Synthesis of fluorescent synthetic peptides capable of conjugation to bioanalytes (nucleic acids, antibodies).

Related Experiment Videos

  • Chemical release of peptides post-assay (hybridization or binding).
  • Detection using a two-step capillary isoelectric focusing (cIEF) method in coated capillaries with salt mobilization, coupled with laser-induced fluorescence (LIF).

    • Main Results:

    • Achieved high peak efficiencies and good assay reproducibility with the developed two-step cIEF method.
    • Demonstrated picomolar (pM) detection limits, significantly lower than sub-nanomolar (sub-nM) levels of conventional CE-LIF, due to the concentration effect of cIEF.
    • Successfully achieved simultaneous multiple detection of eleven different focusing entities.

    Conclusions:

    • The developed method enables analyte-specific detection using fluorescent synthetic peptides and cIEF-LIF.
    • The technique offers enhanced sensitivity (pM detection limits) and multiplexing capabilities.
    • This approach represents a promising advancement for sensitive bioanalyte detection and quantification.