Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Enzymatic production of a protein or polypeptide
Reexamination Certificate
2006-05-09
2006-05-09
Weber, Jon (Department: 1653)
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Enzymatic production of a protein or polypeptide
Reexamination Certificate
active
07041472
ABSTRACT:
The present invention provides a method for selectively collecting the N-terminal peptide fragments of a protein of interest whether or not the protein of interest is modified on the N-terminus. A method for selectively collecting the N-terminal peptide fragment of a protein, comprising: a protection step (1) of protecting side chain-amino groups of amino acid residues containing side chain-amino groups of a protein of interest to obtain a protected protein protected on the side chain-amino groups; a fragmentation step (2) of cleaving the protected protein into one N-terminal peptide fragment (a) containing the N-terminus of the peptide of interest and one or more of peptide fragments (b) other than the N-terminal peptide fragment (a); and a separation step (3) of separating the N-terminal peptide fragment (a) from the other peptide fragments (b) by selectively eluting the N-terminal peptide fragment (a) based on the difference in their reactivity or affinity to substrate, wherein the selective elution is achieved either by allowing the other peptide fragments (b) to bind to the substrate while allowing the N-terminal peptide fragment (a) to elute, or by allowing the N-terminal peptide fragment (a) to bind to the substrate while allowing the other peptide fragments (b) to elute and subsequently eluting the bound N-terminal peptide fragment (a).
REFERENCES:
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patent: WO 02/095412 (2002-11-01), None
Gygi et al., quantitative analysis of complex protein mixtures using isotope-coded affinity tags, Oct. 1999, Nature Biotechnology, vol. 17, pp. 994-999.
W.J. Henzel et al., “Identifying Proteins from Two-dimensional Gels by Molecular Mass Searching of Peptide Fragments in Protein Sequence Database”, Proceedings of the National Academy of Sciences of the United States of America 90 (1993): pp. 5011-5015.
Peter James et al., “Protein Identification by Mass Profile Fingerprinting”, Biochemical and Biophysical Research Communications 195 (1993): pp. 58-64.
D.F. Hunt et al., “Protein Sequencing by Tandem Mass Spectrometry”, Proceedings of the National Academy of Sciences of the United States of America 83 (1986): pp. 6233-6237.
Ando Eiji
Nakazawa Takashi
Norioka Shigemi
Okamura Taka-aki
Ueyama Norikazu
Mondesi Robert B.
Rader & Fishman & Grauer, PLLC
Shimadzu Corporation
Weber Jon
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