Vtutechnology Products & Services

Productivity through diversity – UNLOCK PICHIA through VALIDOGEN’s Pichia pastoris protein production toolbox


Productivity through diversity – UNLOCK PICHIA through VALIDOGEN’s Pichia pastoris protein production toolbox


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Products & Services | VALIDOGEN GmbH

To learn more about VALIDOGEN’s Pichia pastoris protein production toolbox, please click on the brochure below:

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Supplier Information
Supplier: VALIDOGEN GmbH
Address: A-8074 Grambach, Parkring 18, Austria
Tel: +43 (316) 4009 4000
Fax: +43 (316) 4009 4010
Website: www.validogen.com


UNLOCK PICHIA boosting Pichia Glycoswitch® enhanced glycoprotein production


UNLOCK PICHIA boosting Pichia Glycoswitch® enhanced glycoprotein production


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Products & Services | VALIDOGEN GmbH

To learn more about VALIDOGEN’s powerful yield enhancing Pichia pastoris expression platform known as UNLOCK PICHIA
and Pichia GlycoSwitch®, please click on the brochure below:

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Resources

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Supplier Information
Supplier: VALIDOGEN GmbH
Address: A-8074 Grambach, Parkring 18, Austria
Tel: +43 (316) 4009 4000
Fax: +43 (316) 4009 4010
Website: www.validogen.com


UNLOCK PICHIA – Discover VALIDOGEN’s unique methanol-free Pichia protein production system


UNLOCK PICHIA – Discover VALIDOGEN’s unique methanol-free Pichia protein production system


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Products & Services | VALIDOGEN GmbH

To learn more about VALIDOGEN’s unique methanol-free Pichia protein production system, please click on the brochure below:

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Resources

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Supplier Information
Supplier: VALIDOGEN GmbH
Address: A-8074 Grambach, Parkring 18, Austria
Tel: +43 (316) 4009 4000
Fax: +43 (316) 4009 4010
Website: www.validogen.com


Pichia pastoris Protein Expression Excellence – VALIDOGEN Track Record


Pichia pastoris Protein Expression Excellence – VALIDOGEN Track Record


VTU Technology 150 x 56

Products & Services | VTU Technology GmbH

To learn more about Pichia pastoris Protein Expression Excellence – VALIDOGEN Track Record, please click on the brochure below:

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Supplier Information
Supplier: VTU Technology GmbH
Address: A-8074 Grambach, Parkring 18, Austria
Tel: +43 (316) 4009 4000
Fax: +43 (316) 4009 4010
Website: www.vtu-technology.com


Protein Expression Excellence – Pichia pastoris


Protein Expression Excellence – Pichia pastoris


VTU Technology 150 x 56

Products & Services | VTU Technology GmbH

To learn more about Protein Expression Excellence – Pichia pastoris, please click on the brochure below:

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Resources

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Supplier Information
Supplier: VTU Technology GmbH
Address: A-8074 Grambach, Parkring 18, Austria
Tel: +43 (316) 4009 4000
Fax: +43 (316) 4009 4010
Website: www.vtu-technology.com


Pichia pastoris – Protein Production Excellence


Pichia pastoris – Protein Production Excellence


VTU Technology 150 x 56

Products & Services | VTU Technology GmbH

To learn more about Pichia pastoris – Protein Production Excellence, please click on the brochure below:

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Pichia pastoris – Protein Production Excellence

Resources

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Supplier Information
Supplier: VTU Technology GmbH
Address: A-8074 Grambach, Parkring 18, Austria
Tel: +43 (316) 4009 4000
Fax: +43 (316) 4009 4010
Website: www.vtu-technology.com


Custom Glycoprotein Production in Pichia pastoris


Custom Glycoprotein Production in Pichia pastoris


VTU Technology 150 x 56

Products & Services | VTU Technology GmbH

To learn more about Custom Glycoprotein Production in Pichia pastoris, please click on the brochure below:

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Resources

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Supplier Information
Supplier: VTU Technology GmbH
Address: A-8074 Grambach, Parkring 18, Austria
Tel: +43 (316) 4009 4000
Fax: +43 (316) 4009 4010
Website: www.vtu-technology.com


Pichia production of albumin fusion proteins


Pichia production of albumin fusion proteins


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Overview

The use of VTU´s well-established Pichia expression system facilitates high-yield production of albumins and proteins genetically fused to human serum albumin (HSA) using robust, reproducible and scalable processing methods.

The Pichia platform delivers unprecedented capabilities to produce albumin fusion proteins in commercial quantities for the increasing number of biopharma companies with HSA fusion proteins in their pipeline.

VTU´s Pichia pastoris toolbox allows HSA fusion proteins to be secreted very efficiently (5-20 g/L) resulting in reduced production costs at the same time increasing the profitability of a therapeutic product and maintaining quality.

Background

The pharmaceutical and biotech industry is increasingly involved in development of novel biopharmaceuticals and biobetters based on peptides and small proteins with an intrinsic need for half-life extension.

A proven strategy for extending the half-life of therapeutic proteins is fusion to human serum albumin, which is stable, benign and abundant. This fusion allows for less frequent administration while maintaining efficacy.

Albumin is a well characterized, naturally occurring carrier protein with a particularly long serum half-life, which does not elicit unwanted immunogenicity and, in contrast to other technologies, comes with a well-known mechanism of clearance.

Features

VTU´s Pichia pastoris platform is a high productivity system for the manufacture of recombinant albumins and albumin fusion proteins.

This technology has yielded titers of up to 35 g/L of recombinant albumins, thus constituting the highest values so far reported with respect to recombinant proteins being secreted into the culture supernatant of yeast cultivations.

The use of a special subset of VTU´s exclusive AOX1 promoter variants, enabling production processes without the need for induction with methanol, furnishes double-digit g/L levels of HSA in fermentations with glycerol as the sole carbon source.

Benefits

The suite of exclusive technologies making up VTU´s Pichia production platform stands out for its high-performance expression strains and corresponding processes enabling significantly more cost effective production of customer´s HSA fusion proteins.

Compared with alternative technologies, such as PEGylation, Pichia-produced recombinant albumin fusion requires no post-expression chemical modifications and hence fewer purification steps.

VTU Technology has successfully applied recombinant albumin fusion technology to various proteins and peptides. Albiglutide, a dimer of a recombinant glucagon-like peptide-1 variant fused to human albumin ((GLP-1)-(GLP-1)-HSA) has recently obtained US FDA (Tanzeum) and EMA (Eperzan) approvals. Various other proteins and peptides fused to HSA are in clinical and pre-clinical development.

Yields achieved with the VTU Pichia system for albumins and albumin fusion proteins are summarized below:

Protein Titer secreted [g/L]
Albumins up to 35
HSA-IFNα2a 16
HSA-GCSF 18
GCSF-HSA 20
HSA-Chemokine 5
HSA-(GLP-1) 14
PYY(3-36)-HSA-(GLP-1) 13
HSA-(Exendin-4) 11
PYY(3-36)-HSA-(Exendin-4) 11
Scaffold-HSA-Fusions 10 – 15

Resources

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Supplier Information
Supplier: VTU Technology GmbH
Address: A-8074 Grambach, Parkring 18, Austria
Tel: +43 (316) 4009 4000
Fax: +43 (316) 4009 4010
Website: www.vtu-technology.com


Spotlight on methanol-free protein production


Spotlight on methanol-free protein production


VTU Technology

Products & Services | VTU Technology GmbH

VTU Technology has become the world leader in offering Pichia pastoris PAOX1 controlled production processes with unmatched product yields, accelerated development timelines and highest quality standards.

VTU´s exclusive highly approved first-generation library of synthetic methanol-inducible PAOX1 promoter variants forms the core of the company´s cutting-edge in-house Pichia pastoris toolbox enabling high-level protein production and secretion of 10 to 20 g/L with remarkable peak productivities of 35 g/L.

The VTU library has been enhanced with groundbreaking and unique MeOH-free second-generation PAOX1 promoter variants facilitating strong expression without the need for methanol-induction.

The enhanced platform provides safer and more economically viable production processes, sophisticated high-throughput methods and streamlined procedures.

In a recent case study featuring expression of recombinant bacterial Phytase, this broad and versatile production platform hit the 20 g/L mark in methanol-free protein production.

PAOX1 – methanol-free

VTU´s highly proven Pichia pastoris expression toolbox is characterized by a large number of exclusive AOX1 promoter variants, allowing gene expression to be fine-tuned by selecting the perfect match of the respective promoter variant to a given target gene.

VTU´s 2nd generation AOX1 promoter variants are less susceptible to repression by common carbon sources like glucose and glycerol than the AOX1 wild type promoter or VTU´s 1st generation AOX1 promoter variants facilitating strong expression even with just glycerol or glucose as the sole carbon source. Elaborated fermentation and feeding strategies have been developed by VTU to further boost volumetric productivity by reducing the process time while maintaining final product concentrations clearly outperforming conventional promoter systems based on constitutive expression such as pGAP (Figure 1).

Benefits:

VTU´s unique methanol-free Pichia system offers strong expression along with further advantages such as:

  • Banishing toxic and explosive methanol from production process
  • Using glycerol or glucose as the sole carbon source
  • Significantly reduced heat formation and cooling demand at large scale
  • Reduced process time and cost of goods
  • Tunable AOX1 promoter library to comply with different regulatory properties
  • Boosted yields through fine-tuning of gene expression by increasing genetic diversity

Case Study: Methanol-free PAOX1 promoters used in expression of a recombinant bacterial phytase

The case study clearly demonstrates that while maintaining competitive development timelines the application of a higher number of promoter variants from VTU´s promoter libraries significantly boosted protein production in Pichia pastoris. Yield was boosted through fine-tuning of gene expression by increasing genetic diversity (Figure 2).

Fact and figures:

SET UP:
Expression: secreted

  • Methanol-induced expression using VTU´s 1st generation promoter library
  • Methanol-free expression using VTU´s 2nd generation promoter library

Screening size:

  • Small screening – 400 clones applying 2 PAOX1 promoter variants
  • Extended screening – 6000 clones applying 9 PAOX1 promoter variants

Cultivation:

  • Bioreactor size: 0.75 L
  • Pichia strain cultivation using simple and cheap chemically defined media
  • Antibiotic markers used: None

RESULTS:
The study furnished exceptionally high-yielding phytase secreting strains:

  • Methanol-induced: 22 g/L
  • Methanol-free: 20 g/L
  • 3.5-fold to 4.6-fold enhancement of protein yields from small to extended screening through increasing genetic diversity (increasing VTU´s promoter variants and clone numbers)

Resources

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Figure 1 - Accumulations of recombinant target protein in bioreactors

Figure 1: Accumulations of recombinant target protein in bioreactors comparing:
(a) constitutive expression controlled by the GAP-promoter, (b) typical methanol-induced VTU expression, (c & d) MeOH-free processes both controlled by VTU AOX1 promoter variants but using different feeding strategies

Figure 2: Methanol-induced and methanol-free phytase expression based on a small and an extended screening approach

Figure 2: Methanol-induced and methanol-free phytase expression based on a small and an extended screening approach


Supplier Information
Supplier: VTU Technology GmbH
Address: A-8074 Grambach, Parkring 18, Austria
Tel: +43 (316) 4009 4000
Fax: +43 (316) 4009 4010
Website: www.vtu-technology.com


Pichia pastoris Protein Expression Services – Track Record


Pichia pastoris Protein Expression Services – Track Record


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Products & Services | VTU Technology GmbH

BROCHURE

To learn more about VTU’s Pichia pastoris Protein Expression Services – Track Record please click on the brochure below:

Pichia pastoris Protein Expression Services - Track Record

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Supplier Information
Supplier: VTU Technology GmbH
Address: A-8074 Grambach, Parkring 18, Austria
Tel: +43 (316) 4009 4000
Fax: +43 (316) 4009 4010
Website: www.vtu-technology.com


Pichia pastoris Protein Expression Services – AOX1 MeOH-free


Pichia pastoris Protein Expression Services – AOX1 MeOH-free


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Products & Services | VTU Technology GmbH

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To learn more about VTU’s Pichia pastoris Protein Expression Services – AOX1 MeOH-free please click on the brochure below:

Pichia pastoris Protein Expression Services - AOX1 MeOH-free

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Supplier Information
Supplier: VTU Technology GmbH
Address: A-8074 Grambach, Parkring 18, Austria
Tel: +43 (316) 4009 4000
Fax: +43 (316) 4009 4010
Website: www.vtu-technology.com


Pichia pastoris Protein Expression Excellence


Pichia pastoris Protein Expression Excellence


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Products & Services | VTU Technology GmbH

BROCHURE

To learn more about VTU’s Pichia pastoris Protein Expression Excellence please click on the brochure below:

Pichia pastoris Protein Expression Excellence

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Supplier Information
Supplier: VTU Technology GmbH
Address: A-8074 Grambach, Parkring 18, Austria
Tel: +43 (316) 4009 4000
Fax: +43 (316) 4009 4010
Website: www.vtu-technology.com


Pichia Pastoris Protein Expression Services


Pichia Pastoris Protein Expression Services


VTU Technology

Products & Services | VTU Technology GmbH

BROCHURE

To learn more about VTU’s Pichia Pastoris Protein Expression Services please click on the brochure below:

Pichia Pastoris Protein Expression Services

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Supplier Information
Supplier: VTU Technology GmbH
Address: A-8074 Grambach, Parkring 18, Austria
Tel: +43 (316) 4009 4000
Fax: +43 (316) 4009 4010
Website: www.vtu-technology.com


Broadest Pichia pastoris protein expression platform


Broadest Pichia pastoris protein expression platform


VTU Technology

Products & Services | VTU Technology GmbH

Overview

VTU Technology’s Pichia pastoris system provides one of the most powerful and multifaceted recombinant protein expression and production technology platforms holding world productivity records for Pichia-derived recombinant proteins.

Since its foundation 2008, VTU has continuously optimized and expanded its Pichia technology to form an expression toolbox of unparalleled versatility and flexibility for the fast track generation of high performance Pichia pastoris protein production strains and economically viable protein production processes for biopharmaceuticals and other proteins.

Technology base

VTU’s technology suite is based on a cutting-edge Pichia pastoris protein production platform based on its exclusive 1st and 2nd generation AOX1 promoter libraries. These enable sensitive fine-tuning of gene expression by selecting the perfect match out of a huge possible range of promoter/target gene combinations.

The company´s AOX1 promoter libraries control the expression of target genes under two applicable cultivation regimes:

  • VTU´s 1st generation AOX1 promoter library enables unprecedented yields of target protein of more than 20 g/L by secretion into the culture medium upon methanol (MeOH)induction.
  • VTU´s 2nd generation methanol-free AOX1 promoter library functions by derepression even with just glycerol or glucose as the sole carbon source. While retaining high expression levels of up to 15 g/L the use of toxic and explosive methanol as a substrate is completely obviated. Additional major advantages of this new breakthrough technology are reduced oxygen consumption and therefore significantly reduced heat production and cooling effort in bioreactor cultivations as well as a significant potential to reduce process time and cost of goods.

The versatility and effectiveness of VTU´s Pichia system is further underlined by a set of proprietary expression enhancing helper factors, several platform strains with different genetic backgrounds, elaborated cloning and transformation protocols, a high-throughput micro-scale screening and cultivation regime and effective fermentation protocols for maximization of product yield and overall process performance.

Capacities for cultivation and analysis of up to 25.000 clones per week in 96-deep well plates enable the development of customized high performance expression strains with unmatched product yields in minimum time.

With the addition of RCT’s Pichia GlycoSwitch® system for homogeneous N-glycosylation, the VTU portfolio also encompassesa new dimension of recombinant protein expression, creating a high performance production platform for recombinant homogeneous glycoproteins with Man5 and other human-like glycans.

Advantages

VTU´s Pichia pastoris expression system is ideally suited for high-level expression of recombinant proteins for therapeutic or industrial applications, offering many advantages.

These include:

  • A broader platform offering manifold expression and production strategies to meet highly specific expression needs
  • High production levels for a wide range of biologics and other proteins with yields of more than 20 g/L of secreted target protein from MeOH induced processes
  • Up to 15 g/L of secreted target protein from 2nd generation MeOH-free processes that use glycerol or glucose as the sole carbon source, with significantly less heat formation and cooling demand furnishing safe and economically viable production processes
  • Outstanding recombinant glycoprotein product yields and controlled GlcNAc2Man5-pattern or generation of other human-like glycoforms
  • Tight development timelines
  • Competitive production processes

Services

VTU offers a suite of services including:

  • High-speed Pichia pastoris expression strain development
  • Fermentation process development and optimization
  • Downstream and analytical process development
  • Small-scale production of purified non-GMP proteins
  • Sound technology & data transfer to customers
  • Out licensing opportunities for:
    • Human serum albumin and Human serum transferrin
    • HSA-fusion proteins
    • Cell culture proteins

Resources

Click on VTU Pichia pastoris protein expression platform for other information.
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Supplier Information
Supplier: VTU Technology GmbH
Address: A-8074 Grambach, Parkring 18, Austria
Tel: +43 (316) 4009 4000
Fax: +43 (316) 4009 4010
Website: www.vtu-technology.com