What is Peptide Synthesis by Native Chemical Ligation
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What is Peptide Synthesis by Native Chemical Ligation?
Peptide Synthesis by Native Chemical Ligation Since the pioneering days of Bruce Merrifield in solid phase peptide synthesis, in the early 60's, the field of peptide synthesis has considerably evolved; from TBOC to FMOC chemistries, the peptide chemist is now able to obtain fairly long peptides by straight chemical synthesis, being able to attain fragments in the 150 residues in length. However not every peptide is amenable to be assembled by direct chemical synthesis. The idea of being able to ligate native peptides arose from the work of Stephen Kent in 1994 and James Tam and his colleagues in 1995.
The concept lies on being able to use unique functionalities at both the carboxy and amino end of two fully deprotected peptides (native peptides) . In the case of Kent's method, he makes use of an N-terminal Cys that functiosn as the inside thioesther capture, in effect forming anew thioesther via transesterification; the covalent S-acyl intermediate goes tru an S- to N- acyl migration forming a Peptide bond via five membered ring intermediate.
Tam's method was coined chemoselective capture activation ligation and took place via an azothiolidine intermediate. In any event all ligation methods are able to bring the reactive sites of the two segments in close proximity, resulting in intramolecular acyl transfer to establish the peptide bond at the ligation site; that is to say the design brings close to each other the only two possible reactive sites among the 2 peptide fragments, a very ingenious design indeed.
Most ligation procedures take place under aqueous conditions at pH 7-8, in the presence of 6-7M guanidinium chloride; in order to prevent, N,S-bisacylated product and disulfide formation, reducing conditions are recommended (thiophenol addition) In summary we have described briefly the potential benefits of native chemical ligation; whereas a number of long Peptides/proteins, such us IL8 has been accomplished it must be said, that considerable effort and expertise may be needed to master the methodologies for its efficient use.
An excellent review of chemical ligation is given below:
* J.P.Tam, Q.Yu, Z.Miao, Biopolymers 1999, 51, 311
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Add the latest Google news results for your topic, right on your lens. Updates automatically.About BSI
Founded in 1984, Bio-Synthesis, Inc. was first known as OCS Laboratories. It was the first producer of commercially available synthetic DNA and, in 1985, became a producer of synthetic peptides. In 1989, OCS incorporated as Bio-Synthesis, Inc. and moved its laboratories to Lewisville, Texas. Today, BSI occupies 10,000 square feet of modern laboratory space (completed in 1995) located 20 miles from downtown Dallas and 10 miles from D/FW airport. Among the surrounding academic institutions are: University of Texas Southwestern Medical School (12 miles), University of North Texas (10 miles), and University of North Texas Health Science Center (25 miles). These universities provide excellent library resources. Since its inception, BSI has steadily expanded its product lines and services to meet the growing needs of the molecular biology community. It has maintained its position as an aggressive, innovative company in a highly competitive marketplace without sacrificing quality. In the beginning, our primary emphasis was on synthesizing high quality DNA primers and linkers, which were the initial uses of oligos. Today, newer technologies, such as gene construction, PCR, mutagenesis, combinatorial libraries, dye/adduct labeling, DNA microarrays, peptide-nucleic acid chimeras, etc. have challenged the molecular biology field. In response, BSI has branched into several related areas including DNA paternity testing, DNA HLA typing, PNA synthesis, genomic sequencing, fluorescence-based genotyping, custom organic synthesis and other molecular biology based applications.
The Research and Development Division comprises a number of Ph.D. and M.D. professionals with backgrounds in molecular biology, immunology, nucleic acids, peptide synthesis, general organic chemistry, and automated amino acid sequencing. The Molecular Biology Division is set up to handle most modern experimental procedures; the Immunology Division specializes in immunodiagnostic-based reagents.
Not only has BSI continued to provide quality DNA products and services for the research community, but it has also become a world leader in providing custom peptide products and services. Using state-of-the-art solid phase peptide chemistries, BSI provides high quality custom peptides, performs carrier conjugations, antipeptide antibody production, antigenic peptide design, synthesis of long and/or modified peptides, MALDI TOF analysis, contract research, consultation services and more.
Biomedical researchers worldwide in universities, biotech companies, private clinics, and government agencies use products from Bio-Synthesis, Inc. in studies ranging from PCR diagnostics to cancer research and the Human Genome Project. Contact us to discover how BSI can customize products/services to suit your research requirements.
For more detail please visit our About Us
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Address: 612 E. Main street
City: Lewisville
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Phone Number: 972-420-8505
Custom Peptides
As a leading supplier of genomic and proteomic research products, BSI is able to offer its customers integrated solutions for custom DNA synthesis, custom RNA synthesis, custom polyclonal antibody production, custom peptide synthesis, molecular diagnostic detection kits, DNA Real-Time PCR products, DNA microarray services and a wide range of custom DNA-based diagnostic kits and consumables.
The proteomic division is experienced in the design of peptides, both for antibody production and bioactive peptide synthesis, e.g cosmetic peptides. While the custom peptide synthesis division has the ability to produce thousands of modified or non-modified peptides per day for research applications, BSI also provides diagnostic/therapeutic GMP custom peptide production. Synthesis of custom peptides results in quantities ranging from milligrams to multi-kilograms. The staff in the custom antibody division provide expert support in the selection of peptide antigens for target antibody production and offer convenient custom-bundled packages with a wide range of host animals. In addition, peptide, antibody and protein arrays are also amongst its expertise.
BSI is a U.S. based company whose primary emphasis is on the synthesis of high quality custom products for a variety of applications and a full-service contract manufacturing organization (CMO) delivering research, development and production services for cGMP clinical batch manufacturing.
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