Single Protein Expression in Nicotiana

Free 

Description of service

Expression of vaccine antigens in plant-based systems. The Agrobacterium tumefaciens-based system is a robust platform to either rapidly produce up to mg-amounts of recombinant proteins or to compare larger numbers of different vaccine constructs in a screening approach preferentially by transient expression in N. benthamiana plants.

Dependent on the state of development and the focus of the corresponding project, the user may either request one of the following scenarios:

  1. Small scale expression of a larger number (up to 20) of constructs to compare yields, integrity in crude preparations.

  2. Small to medium scale expression (and purification) of a small number (up to 5) of constructs to compare yields, integrity and functionality.

  3. Medium scale expression of one defined vaccine antigen for further studies.

 

Or, may request a combination of the three performed in succession.

Affinity-tags will be used to facilitate easy purification and detection. Depending on the user projects future perspectives, the expression screening will include plant suspension cells and/or initiation of stable plant generation with respect to future manufacturing processes.

Timeline 

2-4 months depending on the complexity of the projects.

Contact 

Holder Spiegel

holger.spiegel@ime.fraunhofer.de

0049241608512461

Related services

The service "Expression of vaccine antigens in plant-based systems" can be requested as a standalone service, or in the context of the "Cross-platform screening and optimization service" which enabled the user to have vaccine antigens expressed in several different platforms with the aim to compare yields, integrity and (if possible) functionality of the protein when produced in different systems.

Once recombinant vaccine antigens produced by this service are available, they could be forwarded to:

Services do NOT include

Costs for shipment of samples will be the responsibility of the User. Costs for synthetic genes will be covered only for one gene (Scenario C) for each user. In case several constructs or construct variants (Scenario A or B) need to be tested, the additional genes have to be supplied by the user.

Possible Output

Scenario A:

Data on expression level, and integrity of the different variants. Recognition by user-supplied antigen-specific mAbs/or sera.

Scenario B:

Data on expression level, and integrity of the different variants, Recognition by user-supplied antigen-specific mAbs/or sera, IMAC-purified proteins in the µg-range (in case of successful expression).

Scenario C:

Data on expression level, and integrity of the different variants, Recognition by user-supplied antigen-specific mAbs/or sera, IMAC-purified proteins in the mg-range (in case of successful expression).

Sample Requirements - input of users

There are different requirements for the different scenarios:

  1. Small scale expression of a larger number (up to 20) of constructs.
    Report featuring all relevant previous data on the vaccine antigen including data from recombinant expression performed by the user or co-workers. Definition of the AA-sequences. Synthetic genes encoding the desired protein sequences. Detection antibodies (if available).

  2. Small to medium scale expression (and purification) of a small number (up to 5) of constructs.
    Report featuring all relevant previous data on the vaccine antigen including data from recombinant expression performed by the user or co-workers. Definition of the AA-sequences. Synthetic genes encoding the desired protein sequences. Detection antibodies (if available). Specification of potential assays to address functionality (if applicable). Specification of requirements (purity, concentration, buffer composition) to perform functionality assays.

  3. Medium scale expression of one defined vaccine antigen.
    Report featuring all relevant previous data on the vaccine antigen including data from recombinant expression performed by the user or co-workers. Definition of the AA-sequences. Detection antibodies (if available). Specification of potential assays to address functionality (if applicable). Specification of requirements (purity, concentration, buffer composition) to perform functionality assays.

Lead Scientist

Holder Spiegel

Publications 

  •  ‘A Plant-Based Transient Expression System for the Rapid Production of Malaria Vaccine Candidates’ (DOI: 10.1007/978-1-4939-3389-1_39)

  • ‘Application of a Scalable Plant Transient Gene Expression Platform for Malaria Vaccine Development’ (DOI: 10.3389/fpls.2015.01169)

  • ‘From Gene to Harvest: insights into upstream process development for the GMP production of a monoclonal antibody in transgenic tobacco plants’ (DOI: 10.1111/pbi.12438)