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Efficient production of single-chain Fv antibody possessing rare codon linkers in fed-batch fermentation

Kumada, Yoichi,Sakan, Yoshinobu,Kajihara, Hideyuki,Kihara, Mana,Kikuchi, Yasufumi,Yamaji, Hideki,Seong, Gi Hun,Katoh, Shigeo Elsevier 2009 Journal of bioscience and bioengineering Vol.107 No.1
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AbstractSingle-chain Fv antibody (scFv) having 2 types of polypeptide linkers with or without rare codons, namely scFv (G4S)3R and scFv No.10 (with rare codons) and scFv (G4S)3 and scFv No.10NR (without rare codon), were expressed under controllable conditions in batch and fed-batch fermentation, in order to compare volumetric productivity and specific productivity levels of scFvs as a soluble form. In batch fermentation, volumetric productivity levels of scFv (G4S)3R and scFv No.10, namely scFvs having the rare codon linkers were 3–5 times higher than those of scFvs that had linkers without the rare codon. In fed-batch fermentation controlled by an exponential feeding system, the cell concentrations of the transformants increased with similar specific growth rates (0.1 h−1), while the specific productivity levels of scFvs with the rare codon linkers were 1.6 times higher than those of scFvs without the rare codon linkers. These results indicate that the presence of several rare codons in the gene of a polypeptide linker increases soluble amount of scFvs. This might be caused by a temporary decrease in translation speed at the position of the polypeptide linker allowing time for the folding of the VH domain and avoiding unfavorable interactions between amino acid residues at the unfolded VH and VL domains. Higher specific productivity levels of both scFv No. 10 and scFv No. 10NR than those of scFv (G4S)3R and (G4S)3 might be caused by difference in stability of the polypeptide linkers on the basis of amino acid sequences. Thus, the rare codon linkers tested in this study will be considerably useful for large-scale production of soluble and active scFvs in fed-batch or continuous fermentations, in which high cell activity can be maintained.
2
Ester기를 갖는 새로운 주쇄형 액정 coumarin 화합물의 합성 및 특성분석

이종백 ( Jong Back Lee ),강병철 ( Byung Chul Kang ),이강춘 ( Gang Choon Lee ),이동진 ( Dong Jin Lee ),( Kihara Hideyuki ) 한국고무학회 2009 엘라스토머 및 콤포지트 Vol.44 No.4
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  ScienceON
  
  KISS
Benzyl 4-Hydroxybenzoate와 1,6-Dibromohexane을 반응시켜 4-(6-bromohexyloxy)benzoic acid를 합성하였다. 이것을 하이드로퀴논과 반응시켜 브롬 그룹을 갖는 새로운 mesogenic ester를 합성하였다. Mesogenic ester를 자외선에 민감한 coumarin과 반응시켜 새로운 형태의 광응답성 coumarin 화합물을 합성 하였다. 합성된 화합물의 구조는 FT-IR 및 1H-NMR에 의해 확인 하였으며, 그들의 열적 상전이온도 및 안정성들은 DSC, 편광현미경 및 X-선 회절에 의해 조사 하였다. 한편 합성된 새로운 화합물은 광학 편광 현미경 관찰에 의해 스멕틱 및 네마틱 구조를 갖는 쌍방성 액정성을 보였다. 4-(6-bromohexyloxy)benzoic acid was synthesized from benzyl 4-Hydroxybenzoate and 1,6-dibromohexane. It was reacted with hydroquinone to obtain a new mesogenic ester having an bromine group. One kind of new photoresponsive coumarin compound was prepared by the reaction of mesogenic ester with coumarin sensitive to the ultraviolet. Structures of the compound were identified by FT-IR and 1H-NMR spectroscopies. Their phase transition temperatures and thermal stability were also investigated by differential scanning calorimetry (DSC), polarized optical microscopy (POM) and x-ray diffraction analysis. From optical polarizing microscopy, the prepared compound was found to show enantiotropic liquid crystallinity with smectic and nematic textures.