Improving the Oral Efficacy of Recombinant Granulocyte Colony-Stimulating Factor and Transferrin Fusion Protein by Spacer Optimization

Bai, Yun; Shen, Wei‐Chiang

doi:10.1007/s11095-006-9059-5

Cited by 113 publications

(85 citation statements)

References 32 publications

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“…PEGylation, [7] acylation, [8] or glycosylation [9] ), fusion with or binding to plasma proteins (e.g. albumin, [10] transferrin, [11] or Fc fragments [12] ) has yielded numerous clinical protein candidates with improved PK/PD profiles. The binding of a polypeptide drug (either covalently or through a tightly binding carrier) to long-lived plasma proteins in the bloodstream enables tight control over targeted drug levels with improved exposure and distribution in the circulation that may potentially lead to lower doses, decreased side effects, and enhanced predictability.…”

mentioning

confidence: 99%

Conjugation of ATIII‐Binding Pentasaccharides to Extend the Half‐Life of Proteins: Long‐Acting Insulin

Kort¹,

Gianotten²,

Wisse³

et al. 2008

ChemMedChem

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mentioning

confidence: 99%

Conjugation of ATIII‐Binding Pentasaccharides to Extend the Half‐Life of Proteins: Long‐Acting Insulin

Kort¹,

Gianotten²,

Wisse³

et al. 2008

ChemMedChem

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“…Such surprising results confused us after at least five times determination. Bai & Shen (2006) indicated that the loss of activity of one moiety is not due to a simple steric hindrance, but rather due to the alteration of the intrinsic conformation of domain in the fusion protein, and such conclusions may offered an explanation for our experimental results.…”

Section: Discussionmentioning

confidence: 56%

In-fusion expression and characterization of β-xylanase and β-1,3-1,4-glucanase in Pichia pastoris

Qiao

Cao

2012

Biologia

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Two chimeric genes, XynA-Bs-Glu-1 and XynA-Bs-Glu-2, encoding Aspergillus sulphureus β-xylanase (XynA, 26 kDa) and Bacillus subtilis β-1,3-1,4-glucanase (Bs-Glu, 30 kDa), were constructed via in-fusion by different linkers and expressed successfully in Pichia pastoris. The fusion protein (50 kDa) exhibited both β-xylanase and β-1,3-1,4-glucanase activities. Compared with parental enzymes, the moiety activities were decreased in fermentation supernatants. Parental XynA and Bs-Glu were superior to corresponding moieties in each fusion enzymes because of lower Km and higher kcat. Despite some variations, common optima were generally 50• C and pH 3.4 for the XynA moiety and parent, and 40• C and pH 6.4 for the Bs-Glu counterparts. Thus, the fusion enzyme XynA-Bs-Glu-1 and XynA-Bs-Glu-2 were bifunctional.

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“…The linker displayed α-helical conformation, which was stabilized by the Glu − -Lys+ salt bridges within segments. They had already been applied to the construction of many recombinant fusion proteins [Amet et al, 2009;Bai and Shen, 2006].…”

Section: Discussionmentioning

confidence: 99%

Influences of Various Peptide Linkers on the Thermotoga maritima MSB8 Nitrilase Displayed on the Spore Surface of Bacillus subtilis

Chen

et al. 2017

Microb Physiol

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In the present study, fusion genes composed of Thermotoga maritima MSB8 nitrilase and Bacillus subtilis 168 outer coat protein CotG were constructed with various peptide linkers and displayed on B. subtilis DB 403 spores. The successful display of CotG-nit fusion proteins on the spore surface of B. subtilis was verified by Western blot analysis and activity measurement. It was demonstrated that the fusion with linker GGGGSEAAAKGGGGS presented the highest thermal and pH stability, which is 2.67- and 1.9-fold of the fusion without linker. In addition, fusion with flexible linker (GGGGS)₃ demonstrated better thermal and pH stability than fusions with linkers GGGGS and (GGGGS)₂. Fusion with rigid linker (EAAAK) demonstrated better thermal stability than fusions with linkers (EAAAK)₂ and (EAAAK)₃. Fusions with linker (EAAAK)₂ demonstrated better pH stability than fusions with linkers (EAAAK) and (EAAAK)₃. In the presence of 1 mM dithiothreitol, 1% (v/v) sodium dodecyl sulfate, and 20% (v/v) ethanol, the optimal linkers of the fusions were MGSSSN, GGGGSEAAAKGGGGS, and (GGGGS)₃, respectively. In summary, our results showed that optimizing the peptide linkers with different type, length, and amino acid composition of the fusion proteins would be an efficient way to maintain the stability of fusion proteins and thus improve the nitrilase display efficiency, which could provide an effective method for rational design peptide linkers of displayed nitrilase on B. subtilis.

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Improving the Oral Efficacy of Recombinant Granulocyte Colony-Stimulating Factor and Transferrin Fusion Protein by Spacer Optimization

Abstract: The insertion of a linker peptide, such as the helix linker H4-2, between G-CSF and Tf domains in the recombinant fusion protein can improve significantly both in vitro and in vivo myelopoietic activity over the non-linker fusion protein.

Cited by 113 publications

References 32 publications

Conjugation of ATIII‐Binding Pentasaccharides to Extend the Half‐Life of Proteins: Long‐Acting Insulin

Conjugation of ATIII‐Binding Pentasaccharides to Extend the Half‐Life of Proteins: Long‐Acting Insulin

In-fusion expression and characterization of β-xylanase and β-1,3-1,4-glucanase in Pichia pastoris

Influences of Various Peptide Linkers on the <b><i>Thermotoga maritima</i></b> MSB8 Nitrilase Displayed on the Spore Surface of <b><i>Bacillus subtilis</i></b>

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