Title:
ポリペプチドの配列バリアントの決定方法
Document Type and Number:
Japanese Patent JP5903051
Kind Code:
B2
Abstract:
The invention is directed to a method for determining amino acid sequence mutations in a produced polypeptide, comprising the following steps of a) providing a sample of a produced polypeptide, b) incubating the polypeptide in the sample with a protease, c) performing a two dimensional analysis using reversed phase chromatography coupled with a high resolution mass spectroscopy (FT-ICR/FT-orbitrap) and MS/MS analysis of the amino acid sequence fragments of the peptides, d) data evaluation by comparing the LC-MS data sets obtained for the samples side by side with the data set of a reference sample, by searching for differences in the signal intensities at given retention times and by evaluation of differential signals with respect to amino acid sequence mutations. The reference sample for data evaluation (d) can be either a well characterized standard or one of the samples to be analyzed.
Inventors:
Cole Hands
Regra yoerk thomas
Wiesov Frank
Zeck Ann
Regra yoerk thomas
Wiesov Frank
Zeck Ann
Application Number:
JP2012552430A
Publication Date:
April 13, 2016
Filing Date:
February 16, 2011
Export Citation:
Assignee:
F. HOFFMANN-LA ROCHE AKTIENGESELLSCHAFT
International Classes:
G01N27/62; C12Q1/37; G01N33/68
Domestic Patent References:
| JP2006515987A | ||||
| JP2004503792A | ||||
| JP2005536714A | ||||
| JP2005510732A |
Other References:
SREBALUS BARNE,C.A. 外1名,“Applications of mass spectrometry for the structural characterization of recombinant protein pharmaceuticals”,Mass Spectrometry Reviews,2007年4月4日,Volume 26,Issue 3,Pages 370-388
YU,X.C.外5名,“Identification of Codon-Specific Serine to Asparagine Mistranslation in Recombinant Monoclonal Antibodies by High-Resolution Mass Spectrometry”,Analytical Chemistry,2009年10月23日,Volume 81,Number 22,Pages 9282-9290
YU,X.C.外5名,“Identification of Codon-Specific Serine to Asparagine Mistranslation in Recombinant Monoclonal Antibodies by High-Resolution Mass Spectrometry”,Analytical Chemistry,2009年10月23日,Volume 81,Number 22,Pages 9282-9290
Attorney, Agent or Firm:
Hatsushi Shimizu
Masao Haruna
Hirotaka Yamaguchi
Toshi Gobe
Ryuichi Inoue
Toshimitsu Sato
Koichi Niimi
Tomohiko Kobayashi
Masato Ozeki
Yoshihiro Igarashi
Kazuya Kawamoto
Masao Haruna
Hirotaka Yamaguchi
Toshi Gobe
Ryuichi Inoue
Toshimitsu Sato
Koichi Niimi
Tomohiko Kobayashi
Masato Ozeki
Yoshihiro Igarashi
Kazuya Kawamoto