Optimization of a fusion protein expression system using human cell lines to create a practical immunoassay reagent

Kounosuke Hayashi, Yusuke Tomozoe, Kenji Nagai, Kyoichi Matsuba, Masayuki Mitsumori, Yoshiyuki Hiraishi, Toshiharu Matsumura, Hiroshi Ueda, Noriho Kamiya

Research output: Contribution to journalArticle

2 Citations (Scopus)

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 Staphylococcus aureus Protein A and Streptococcus 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.

Original languageEnglish
Pages (from-to)38-42
Number of pages5
Journalkagaku kogaku ronbunshu
Volume41
Issue number1
DOIs
Publication statusPublished - Jan 20 2015

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Fusion reactions
Cells
Proteins
Enzymes
Antibodies
DNA sequences
Staphylococcal Protein A
Labeling
Alkaline Phosphatase
Suspensions
Carrier Proteins
Plasmids
Phosphatases
indolepropanol phosphate

All Science Journal Classification (ASJC) codes

  • Chemistry(all)
  • Chemical Engineering(all)

Cite this

Optimization of a fusion protein expression system using human cell lines to create a practical immunoassay reagent. / Hayashi, Kounosuke; Tomozoe, Yusuke; Nagai, Kenji; Matsuba, Kyoichi; Mitsumori, Masayuki; Hiraishi, Yoshiyuki; Matsumura, Toshiharu; Ueda, Hiroshi; Kamiya, Noriho.

In: kagaku kogaku ronbunshu, Vol. 41, No. 1, 20.01.2015, p. 38-42.

Research output: Contribution to journalArticle

Hayashi, K, Tomozoe, Y, Nagai, K, Matsuba, K, Mitsumori, M, Hiraishi, Y, Matsumura, T, Ueda, H & Kamiya, N 2015, 'Optimization of a fusion protein expression system using human cell lines to create a practical immunoassay reagent', kagaku kogaku ronbunshu, vol. 41, no. 1, pp. 38-42. https://doi.org/10.1252/kakoronbunshu.41.38
Hayashi, Kounosuke ; Tomozoe, Yusuke ; Nagai, Kenji ; Matsuba, Kyoichi ; Mitsumori, Masayuki ; Hiraishi, Yoshiyuki ; Matsumura, Toshiharu ; Ueda, Hiroshi ; Kamiya, Noriho. / Optimization of a fusion protein expression system using human cell lines to create a practical immunoassay reagent. In: kagaku kogaku ronbunshu. 2015 ; Vol. 41, No. 1. pp. 38-42.
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