Chemistry: analytical and immunological testing – Peptide – protein or amino acid – Amino acid or sequencing procedure
Reexamination Certificate
2011-02-01
2011-02-01
Gakh, Yelena G (Department: 1797)
Chemistry: analytical and immunological testing
Peptide, protein or amino acid
Amino acid or sequencing procedure
C436S086000, C436S090000, C435S023000, C530S345000
Reexamination Certificate
active
07879616
ABSTRACT:
The present invention provides a method for analyzing the C-terminal amino acid sequence of a peptide by using a reaction for successively releasing the C-terminal amino acids of the peptide, which method can suppress, when successively releasing the C-terminal amino acids of a peptide of long amino acid length, such a undesirable side reaction as cleavage of peptide bond in the intermediate position of the peptide and can carry out the chemical treatment thereof under widely applicable conditions; In the method, a dry sample of a peptide with long amino acid length is beforehand subjected to an N-acylation treatment; by using a reaction reagent where an alkanoic acid anhydride is combined with a small amount of a perfluoroalkanoic acid, successive release of C-terminal amino acids is conducted under mild conditions; a hydrolysis treatment is applied; then, selective fragmentization at site of arginine residue is performed by digestion by trypsin; thereafter, decreases in molecular weight are measured for the C-terminal side fragments derived from a series of reaction products with use of a MALDI-TOF-MS apparatus; thereby, the C-terminal amino acid sequence of the peptide sample is identified.
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Kamijo Kenichi
Miyazaki Kenji
Torii Hiroaki
Tsugita Akira
Gakh Yelena G
NEC Corporation
Scully , Scott, Murphy & Presser, P.C.
Xu Robert
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