Description of service

SPR-based molecular interaction analysis. In the context of this service IME will develop and perform assays that can be used to either:

  1. Characterize the interaction between biomolecules (e.g. protein and natural ligand, protein and conformational mAb) on quantitative and qualitative level.

  2. Characterize, and quantify antibody responses in serum samples (before, during or after immunization studies).

In the context of vaccine development and vaccine manufacturing these assays can provide a high value. Using so called calibration-free concentration analysis (CFCA) it is usually possible to address the total concentration of vaccine antigen specific antibodies in serum samples. Analysis interaction of vaccine antigens with natural ligand or antibodies can be used to investigate the activity and homogeneity of the protein and can be implemented during vaccine candidate development, process development and quality control.

Timeline 

2-4 weeks, upon the availability of all required samples and reagents.  

 4-6 weeks for modification of existing or development of new assay formats

Contact 

Holger Spiegel

holger.spiegel@ime.fraunhofer.de

0049241608512461

Services do NOT include

Costs for shipment of samples will be the responsibility of the User. Recombinant proteins required as interaction partner (apart from standard capture ligands like protein A/G/L or human, mouse or rabbit antibody specific ligands).

Possible Output

Depending on the character of the analysis:

  1. Absolute or relative binding activities

  2. Thermodynamic interaction constants (kon, koff, KD, ΔH and ΔS)

  3. Stoichiometry of interaction

  4. Absolute concentrations of antigen specific antibodies

  5. Other parameters

Sample Requirements - input of users

The specification of sample requirements will vary from project to project.

In general recombinant protein samples should have a high degree of purity (>75%).

High concentrations are helpful (1mg/ml). The minimum amounts strongly depend on the stoichiometry of the interaction, the affinity of the interaction as well as the molecular weights of the interaction partners. As a rule of thumb at least 100µg of the interaction partners should be available.

Sequences, biophysical properties, buffer compositions and further information on the ligands have to be provided by the user, together with a documentation demonstrating the state of purity and integrity (if applicable).

For the characterization of immune Ig, purified antibody preparations (e.g. Protein G or Protein A) are preferred. In case of human sera special requirements may apply.

Lead Scientist

Holger Spiegel

SPR-analytics (kinetics, binding parameters, CFCA)

Free 

Related services

This service could be utilized to analyse samples derived in the context of other services offered by TRANSVAC2, including the recombinant vaccine antigens provided by the "Cross platform screening and optimization service", the "Pre-clinical GLP production service" or even the "GMP production service". Furthermore serum samples or immune Ig preparations generated in the context of immunogenicity and challenge studies performed under the umbrella of the "Animal Models" could be analysed.

Publications

  • Analysis of a multi-component multi-stage malaria vaccine candidate-tackling the cocktail challenge (DOI: 10.1371/journal.pone.0131456)

  • Immunization with the Malaria Diversity-Covering Blood-Stage Vaccine Candidate Plasmodium falciparum Apical Membrane Antigen 1 DiCo in Complex with Its Natural Ligand PfRon2 Does Not Improve the In Vitro Efficacy (DOI: 10.3389/fimmu.2017.00743)

  • Detailed functional characterization of glycosylated and nonglycosylated variants of malaria vaccine candidate PfAMA1 produced in Nicotiana benthamiana and analysis of growth inhibitory responses in rabbits. (DOI: 10.1111/pbi.12255)