成果報告書詳細
管理番号20090000000127
タイトル*平成20年度中間年報 「新機能抗体創製技術開発/新機能抗体創製技術開発」 平成18年度-平成21年度 「新機能抗体創製技術開発/高効率な抗体分離精製技術の開発」 平成18年度-平成22年度
公開日2009/12/9
報告書年度2008 - 2008
委託先名財団法人バイオインダストリー協会 独立行政法人産業技術総合研究所
プロジェクト番号P06009
部署名バイオテクノロジー・医療技術開発部 健康グループ
和文要約以下本編抜粋:1.研究開発の内容、成果等 本研究の目標は、以下の技術開発である。(1)膜タンパク質及びその複合体等の機能を有した抗原の系統的な産生技術 創薬標的となりうる産生が困難な膜タンパク質やその複合体等(Gタンパク質共役型受容体(GPCR)等)を、生体内における機能を有した状態で、系統的に産生する技術開発を行う。特に、高い特異性を有する抗体の創製に適した抗原産生技術を開発する。
英文要約Title:Development of New Functional Antibody Technologies (FY2006-FY2008) FY2008 Annual Reort
A)Development of integrated technology to generate antibodies of high specificity
By immunization of these viruses, we obtained specific monoclonal antibodies against
16 membrane proteins in total. We have developed a rapid and efficient method to make
recombinant baculovirus which display part of target proteins. We have prepared 171
recombinant viruses which display marker candidate proteins for various cancers,
diabetes, neuropsychiatric disorder, or cardiovascular diseases.We further established a
monoclonal antibody which can deplete regulatory T cells in vivo, and identified its
specific antigen. We also have developed engineered fragment of tumor-specific
monoclonal antibodies attempting to improve their stability and high tumor specific
accumulation for PET in vivo imaging technology.Using the system we selected 31
cancer specific membrane molecules from gene expression profiles of 101 breast cancer
and 28 molecules from 46 esophageal cancer. In order to establish advanced gene
expression profiling system, we have started to collect clinical samples of 16 different
types of cancer and now obtained more than 150 cases from 8 types of cancer. And we
have analyzed gene expression analysis using exon array and developed exon array
database and data viewer system. Using this system we determined 2 new target
molecules against lung adenocarcinoma. High affinity and low-noise magnetic beads
have been developed to analyze ultra-low abundant target proteins in the cell. High
affinity monoclonal antibodies generated in this project were utilized for this AP-MS
(affinity purification mass-spectrometry) method to detect associating partner proteins
of endogenous target protein complex.
B) Development of High Performance Separation Technologies for Antibodies
As for affinity ligand proteins, more than 750 mutant proteins were produced, and their
IgG-binding characteristics were examined. The developed instrument for
high-through-put analysis was further improved to enable us to measure elution
profiles of IgG from immobilized ligands to be tested simultaneously. As for aggregation
detecting technologies, FTIR spectra of antibody samples were statistically analyzed
using a new method.Silica-based packing for affinity chromatography with the chemical
stability was developed, which has the potentiality to utilize for the purification of
monoclonal antibodies in industrial scale. Thin layer monolith silica gel on the glass
plate and micro size monolith column were developed for ligand evaluation instrument.
Three Chinese hamster ovary (CHO) cell lines producing IgG-like model antibodies
were constructed and adapted to serum-free medium. The kinetic parameters and the
antibody variety were evaluated in small-scale fed-batch culture using constructed
Lectin binding assay. The scale-up culture study for three CHO cell lines was performed.
The biological activities of IgG-like model antibodies produced in industrial scale were
found to be comparable to those of the antibodies prepared in laboratory scale.A simple
and useful method for designing an efficient stepwise elution chromatography process
was proposed. The method was verified by both a small column and a 96-well microplate
based high throughput system.
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