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

Chemoselective one-step purification method for peptides synthesized by the solid-phase technique.

S Funakoshi1, H Fukuda, N Fujii

  • 1Kyoto University, Japan.

Proceedings of the National Academy of Sciences of the United States of America
|August 15, 1991
PubMed
Summary

A new affinity purification method uses a specific reaction between SH and iodoacetamide groups to efficiently purify synthesized peptides. This technique simplifies peptide purification, yielding high-purity results for solid-phase synthesis applications.

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

[Continuous epidural infusion of bupivacaine and buprenorphine for postoperative pain relief].

Masui. The Japanese journal of anesthesiology·1992
Same author

[Sevoflurane anesthesia for asthmatic patients: report of five cases].

Masui. The Japanese journal of anesthesiology·1992
Same author

Occurrence of the nfxB type mutation in clinical isolates of Pseudomonas aeruginosa.

Antimicrobial agents and chemotherapy·1992
Same author

Identification of the binding site of 55kDa tumor necrosis factor receptor by synthetic peptides.

Biochemical and biophysical research communications·1992
Same author

Two reactions are simultaneously catalyzed by a single enzyme: the arginine-dependent simultaneous formation of two products, ethylene and succinate, from 2-oxoglutarate by an enzyme from Pseudomonas syringae.

Biochemical and biophysical research communications·1992
Same author

Molecular cloning in Escherichia coli, expression, and nucleotide sequence of the gene for the ethylene-forming enzyme of Pseudomonas syringae pv. phaseolicola PK2.

Biochemical and biophysical research communications·1992

Area of Science:

  • Biochemistry and Organic Chemistry
  • Peptide Synthesis and Purification

Background:

  • Solid-phase peptide synthesis (SPPS) is a common method for creating peptides.
  • Purification of synthesized peptides can be challenging, impacting downstream applications.
  • Existing methods may lack efficiency or specificity for certain peptide purification needs.

Purpose of the Study:

  • To develop a novel affinity-type purification procedure for solid-phase synthesized peptides.
  • To utilize the specific reaction between thiol (SH) and iodoacetamide groups for peptide purification.
  • To create a robust method applicable to both 9-fluorenylmethoxycarbonyl (Fmoc) and tert-butoxycarbonyl (Boc) based SPPS.

Main Methods:

  • Synthesis of an SH precursor reagent with an acid-labile S-protecting group (pMB).

Related Experiment Videos

  • Attachment of the SH precursor to the peptide-resin via a base-labile linkage during SPPS.
  • Acid treatment to deprotect and cleave the peptide, followed by immobilization on an iodoacetamide-resin column.
  • Base treatment to elute the purified peptide from the affinity column.

    • Main Results:

    • The developed method successfully purified peptides ranging from 18 to 44 amino acids in length.
    • Peptides were obtained in nearly pure form after the four-step affinity purification process.
    • The procedure demonstrated effectiveness with Fmoc-based SPPS and showed potential for Boc-based SPPS.

    Conclusions:

    • The SH-iodoacetamide affinity purification method provides an efficient and specific approach for peptide purification.
    • This technique offers a valuable tool for researchers involved in solid-phase peptide synthesis.
    • The method's adaptability to different SPPS strategies enhances its broad applicability in peptide chemistry.