固相免疫測定を目的とした融合タンパク質試薬の設計とヒト培養細胞による生産プロセスの最適化

Translated title of the contribution: Optimization of a Fusion Protein Expression System using Human Cell Lines to Create a Practical Immunoassay Reagent

林 浩之輔, 友添 祐介, 永井 賢治, 松葉 恭一, 三ツ森 正之, 平石 佳之, 松村 外志張, 上田 宏, 神谷 典穂

Research output: Contribution to journalArticlepeer-review

Abstract

New fusion proteins for immunoassay were developed as an alternative to the conventional chemical linked enzyme-antibody complex. Human chimeric alkaline phosphatase (IPP) was used as the labeling enzyme, and partial domains of <i>Staphylococcus aureus</i> Protein A and <i>Streptococcus</i> Protein G were used as antibody binding protein (PG). The fusion protein composed of IPP and PG was produced using human cell lines because IPP is derived from human enzymes. The expression system was optimized by changing the plasmid vector, reducing the GC content of the DNA sequence, and employing different cell lines including adherent and suspension cells. As result, a 2,600-fold increase in the protein yield per unit of growth medium was achieved. The resultant IPP-PG fusion protein was successfully utilized for immunoassay applications such as western blotting and immunohistochemistry.
Translated title of the contributionOptimization of a Fusion Protein Expression System using Human Cell Lines to Create a Practical Immunoassay Reagent
Original languageJapanese
Pages (from-to)38-42
Number of pages5
JournalKagaku Kogaku Ronbunshu
Volume41
Issue number1
DOIs
Publication statusPublished - 2015

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