Removing boundaries for therapeutic protein production in the filamentous fungus Trichoderma reesei: Reduction of the host protease load

Christopher Landowski, Anne Huuskonen, Ramon Wahl, Ann Westerholm-Parvinen, Benjamin Sommer, Merja Penttilä, Jari Natunen, Christian Ostermeier, Bernhard Helk, Juhani Saarinen, Markku Saloheimo (Corresponding author)

Research output: Contribution to conferenceConference AbstractScientific

Abstract

The filamentous fungus Trichoderma reesei is an important protein production hosts in the enzyme industry. It produces native cellulase and hemicellulase enzymes with extremely high efficiency; the highest enzyme yields exceed 100 g/l. The success in production of mammalian proteins has thus far been much more limited. Antibody Fab fragments and bovine chymosin have been produced at over 100 mg/l levels in earlier work. We report results from a new effort to develop a T. reesei platform for therapeutic protein production. This effort has been much more successful, leading to production levels of several g/l for several therapeutic molecules. The production development of two proteins, full-length IgG antibodies and interferon α2b will be reported. After constructing the first expression strains it became clear that the host proteases formed a major boundary for their production. The secreted T. reesei proteases were characterised with respect to their biochemical properties, their activity against the target proteins and their abundance at mRNA and protein levels. The data obtained was used to make a series of T. reesei strains with multiple protease deletions. The general protease activity in the best deletion strains was ca. 30-fold reduced as compared to the original strain. In the best constructed strains both IgG antibodies and interferon ?2b could be produced at above 7 g/l. Interestingly, the protease deletion strains had a faster growth rate than the original strains. This study shows the excellent potential of T. reesei as a host for therapeutic protein production.
Original languageEnglish
Publication statusPublished - 2015
Event8th Conference on Recombinant Protein Production, RPP8 - Palma de Mallorca, Spain
Duration: 22 Apr 201524 Apr 2015

Conference

Conference8th Conference on Recombinant Protein Production, RPP8
Abbreviated titleRPP8
CountrySpain
CityPalma de Mallorca
Period22/04/1524/04/15

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biopharmaceuticals
Trichoderma reesei
proteinases
fungi
interferons
proteins
antibodies
enzymes
chymosin
endo-1,4-beta-glucanase
industry
therapeutics
cattle

Cite this

Landowski, Christopher ; Huuskonen, Anne ; Wahl, Ramon ; Westerholm-Parvinen, Ann ; Sommer, Benjamin ; Penttilä, Merja ; Natunen, Jari ; Ostermeier, Christian ; Helk, Bernhard ; Saarinen, Juhani ; Saloheimo, Markku. / Removing boundaries for therapeutic protein production in the filamentous fungus Trichoderma reesei : Reduction of the host protease load. Abstract from 8th Conference on Recombinant Protein Production, RPP8, Palma de Mallorca, Spain.
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title = "Removing boundaries for therapeutic protein production in the filamentous fungus Trichoderma reesei: Reduction of the host protease load",
abstract = "The filamentous fungus Trichoderma reesei is an important protein production hosts in the enzyme industry. It produces native cellulase and hemicellulase enzymes with extremely high efficiency; the highest enzyme yields exceed 100 g/l. The success in production of mammalian proteins has thus far been much more limited. Antibody Fab fragments and bovine chymosin have been produced at over 100 mg/l levels in earlier work. We report results from a new effort to develop a T. reesei platform for therapeutic protein production. This effort has been much more successful, leading to production levels of several g/l for several therapeutic molecules. The production development of two proteins, full-length IgG antibodies and interferon α2b will be reported. After constructing the first expression strains it became clear that the host proteases formed a major boundary for their production. The secreted T. reesei proteases were characterised with respect to their biochemical properties, their activity against the target proteins and their abundance at mRNA and protein levels. The data obtained was used to make a series of T. reesei strains with multiple protease deletions. The general protease activity in the best deletion strains was ca. 30-fold reduced as compared to the original strain. In the best constructed strains both IgG antibodies and interferon ?2b could be produced at above 7 g/l. Interestingly, the protease deletion strains had a faster growth rate than the original strains. This study shows the excellent potential of T. reesei as a host for therapeutic protein production.",
author = "Christopher Landowski and Anne Huuskonen and Ramon Wahl and Ann Westerholm-Parvinen and Benjamin Sommer and Merja Penttil{\"a} and Jari Natunen and Christian Ostermeier and Bernhard Helk and Juhani Saarinen and Markku Saloheimo",
note = "CA2: BA3113 CA2: BA311 SDA: SHP: Bioeconomy; 8th Conference on Recombinant Protein Production, RPP8, RPP8 ; Conference date: 22-04-2015 Through 24-04-2015",
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language = "English",

}

Landowski, C, Huuskonen, A, Wahl, R, Westerholm-Parvinen, A, Sommer, B, Penttilä, M, Natunen, J, Ostermeier, C, Helk, B, Saarinen, J & Saloheimo, M 2015, 'Removing boundaries for therapeutic protein production in the filamentous fungus Trichoderma reesei: Reduction of the host protease load', 8th Conference on Recombinant Protein Production, RPP8, Palma de Mallorca, Spain, 22/04/15 - 24/04/15.

Removing boundaries for therapeutic protein production in the filamentous fungus Trichoderma reesei : Reduction of the host protease load. / Landowski, Christopher; Huuskonen, Anne; Wahl, Ramon; Westerholm-Parvinen, Ann; Sommer, Benjamin; Penttilä, Merja; Natunen, Jari; Ostermeier, Christian; Helk, Bernhard; Saarinen, Juhani; Saloheimo, Markku (Corresponding author).

2015. Abstract from 8th Conference on Recombinant Protein Production, RPP8, Palma de Mallorca, Spain.

Research output: Contribution to conferenceConference AbstractScientific

TY - CONF

T1 - Removing boundaries for therapeutic protein production in the filamentous fungus Trichoderma reesei

T2 - Reduction of the host protease load

AU - Landowski, Christopher

AU - Huuskonen, Anne

AU - Wahl, Ramon

AU - Westerholm-Parvinen, Ann

AU - Sommer, Benjamin

AU - Penttilä, Merja

AU - Natunen, Jari

AU - Ostermeier, Christian

AU - Helk, Bernhard

AU - Saarinen, Juhani

AU - Saloheimo, Markku

N1 - CA2: BA3113 CA2: BA311 SDA: SHP: Bioeconomy

PY - 2015

Y1 - 2015

N2 - The filamentous fungus Trichoderma reesei is an important protein production hosts in the enzyme industry. It produces native cellulase and hemicellulase enzymes with extremely high efficiency; the highest enzyme yields exceed 100 g/l. The success in production of mammalian proteins has thus far been much more limited. Antibody Fab fragments and bovine chymosin have been produced at over 100 mg/l levels in earlier work. We report results from a new effort to develop a T. reesei platform for therapeutic protein production. This effort has been much more successful, leading to production levels of several g/l for several therapeutic molecules. The production development of two proteins, full-length IgG antibodies and interferon α2b will be reported. After constructing the first expression strains it became clear that the host proteases formed a major boundary for their production. The secreted T. reesei proteases were characterised with respect to their biochemical properties, their activity against the target proteins and their abundance at mRNA and protein levels. The data obtained was used to make a series of T. reesei strains with multiple protease deletions. The general protease activity in the best deletion strains was ca. 30-fold reduced as compared to the original strain. In the best constructed strains both IgG antibodies and interferon ?2b could be produced at above 7 g/l. Interestingly, the protease deletion strains had a faster growth rate than the original strains. This study shows the excellent potential of T. reesei as a host for therapeutic protein production.

AB - The filamentous fungus Trichoderma reesei is an important protein production hosts in the enzyme industry. It produces native cellulase and hemicellulase enzymes with extremely high efficiency; the highest enzyme yields exceed 100 g/l. The success in production of mammalian proteins has thus far been much more limited. Antibody Fab fragments and bovine chymosin have been produced at over 100 mg/l levels in earlier work. We report results from a new effort to develop a T. reesei platform for therapeutic protein production. This effort has been much more successful, leading to production levels of several g/l for several therapeutic molecules. The production development of two proteins, full-length IgG antibodies and interferon α2b will be reported. After constructing the first expression strains it became clear that the host proteases formed a major boundary for their production. The secreted T. reesei proteases were characterised with respect to their biochemical properties, their activity against the target proteins and their abundance at mRNA and protein levels. The data obtained was used to make a series of T. reesei strains with multiple protease deletions. The general protease activity in the best deletion strains was ca. 30-fold reduced as compared to the original strain. In the best constructed strains both IgG antibodies and interferon ?2b could be produced at above 7 g/l. Interestingly, the protease deletion strains had a faster growth rate than the original strains. This study shows the excellent potential of T. reesei as a host for therapeutic protein production.

M3 - Conference Abstract

ER -