The Biopharmaceutical Lab offers vast experience in bioassays and protein chemistry, as well as a wide range of analytic techniques and lab equipment. It provides solutions in the field of analytical control of biopharmaceuticals, either using pre-established methodologies based on international guides and pharmacopeia, or developing new analytic tools in order to meet and follow bioanalytical strategies.
Our assays follow ICH guidelines and the FDA and EMA regulations.
Created in June 2009 under the direction of Alejandro Ricciardi, the lab was established by the Ministry of Public Health as a national reference laboratory for the control of biopharmaceuticals sold in the Uruguayan market.
To fulfill this task, the Biopharmaceutical Laboratory has highly qualified human resources, equipment and the support of different technological platforms of the Institut Pasteur de Montevideo. Our services are developed under GLP conditions (Certified by LSQA, Enabled by MSP), and in accordance with the directives established by the ICH guidelines, as well as the FDA and EMA agencies.
On biopharmaceuticals, the focus of study of the laboratory, we have developed specific kits and analytical methodologies base on requests from productive sector. Some of the projects that we are working on are:
Development of Methodologies to Quantify Proteins and DNA Contaminant Derivatives of the Host Cell in Recombinant Bio-Pharmaceuticals. Funded by the National Research and Innovation Agency. (Project ALIANZA – Laboratorio Celsius S.A. and IP Montevideo) (2011 – 2012).
Methodological development for quantification of immunogenicity generated by administration of Interferon beta1a in patients, by means of a Bioassay based on cell culture and Real Time PCR. Funded by Laboratorios Clausen S.A. (2010).
Participation in a Multicenter Study for the Determination of the Biological Potency of the First Filgrastim Standard of the United States Pharmacopeia (USP). (2012). (2012)
- HPLC Prominence with DAD, RID and Fluorescence detectors (Shimadzu)
- PA 800 Plus Capillary Electrophoresis (Beckman Coulter)
- Dicroísmo CircularApplied Photophysics (Chiriascan V100)
- Multiskan Spectrum Spectrophotometer and Plate Reader (Thermo Scientific)
- Class II, Type A2 Biological Safety Cabinet (Thermo Scientific)
- CO2Incubator (Thermo Scientific)
- InvertedMicroscope (Nikon)
A) Quality Control of Biopharmaceuticals
- Biological Activity: cell-based bioassays, kinetic assays, and in vivo assays in different species.
- Purity Assays: performed by HPLC, SDS-PAGE, zone and Capillary Electrophoresis, isoelectric focusing or 2D electrophoresis. And either ELISA to quantify protein contaminants or hybridization for DNA contaminants.
- Identification Assays: are done through immunochemistry techniques, peptide mapping, N-glycan profiling.
- And quantification assays through colorimetric and HPLC techniques.
B) Institutional Technological Platform for Biopharmaceutical Comparability Studies
The current regulations and international guidelines establish new and rigorous quality requirements to demonstrate biosimilarity among the innovative products already existent in the market and its possible copies.
These requirements are important in a potential biosimilar development stage, to generate scientific evidence supporting the quality, efficacy and safety of the biosimilar to be as close as possible to the reference product.
The comparability study from which biosimilarity should be inferred consists overall in three steps:
1) Physicochemical and biological quality comparability “in vitro”
Complete characterization of physicochemical, biological and immunohistochemical properties of MBS and its comparison under equal conditions with MBR. Impurity profile.
Study and evaluation of the clinical impact of the existing differences.
2) Non-clinical comparability
It depends on the type of product and the similarity demonstrated in the comparability of Quality. It is evaluated case by case.
In vitro studies: they are generally carried out during the comparability of Quality
In vivo studies of biological activity and toxicity. Non-clinical comparability and finally
3) Clinical comparability
The health authority will define the need to carry out clinical trials
Comparative Pharmacokinetic and Pharmacodynamic Studies
The physicochemical and biological characterization is the analytical founding for the development and comparison of the possible biosimilars, and the amount of possible reduction for non-clinical and clinical comparison studies depends of the success in this first stage.
We have experience in biosimilars head to head physicochemical comparability studies in our Lab, together with other platforms of the Institute, following WHO and EMA international guidelines. Besides the previously described assays, the analytical set for comparability studies include: binding assays, folding assays, characterization and quantification of molecular aggregates, thermal stability, and tertiary structure determination among others.
Technological Transfer to Laboratorio Celsius S.A. (Uruguay) for the Biological Activity Bioessay of Filgrastim, (2009).
Methodological Development for the Evaluation of Immunogenicity for the case of Interferon beta 1a, in patients in treatment. Laboratorios Clausen S.A. (Uruguay), (2010).
Analytical Technological Transfer of Biopharmaceuticals to the Biocertifica Consortium (Chile), (2010).
Development of Methodologies to Quantify Proteins and DNA Contaminant Derivatives of the Host Cell in Recombinant Biopharmaceuticals. Project ALLIANCE financed by ANII and Laboratorio Celsius S.A., (2011 – 2012).
Bioessay Validation Study for Biological Power of Filgrastim for the Eurofarma Laboratory (Brazil), (2012).
Physicochemical Comparability Study between two commercial biopharmaceuticals of Abciximab, Laboratorio Libra S.A., (Uruguay), 2013.
Technological Transfer of Biogen (USA) for the Biological Activity Bioessay of Interferon beta 1a., (2013).
Bioessay Validation Study for Biological Potency of Peg-Filgrastim for the Eurofarma Laboratory (2014).
Participation in the Physicochemical Comparability Studies for the Development of a Biosimilar based on Filgrastim (Fiprima) from the Eurofarma Laboratory, Brazil. First Biosimilar of original production in Latin America authorized by ANVISA, (2011 – 2015).
Technological Transfer to Eurofarma, Brazil, of the analytical methodology for the realization of the Biological Activity based on Cell Culture for Filgrastim and Peg-Filgrastim, (2016).
Participation in the Comparability Studies for the Development of a Biosimilar based on Erythropoietin and Filgrastrim for a Regional Laboratory. (2017 – 2019).
- Turell L, Botti H, Bonilla L, Torres MJ, Schopfer F, Freeman BA, Armas L, Ricciardi A, Alvarez B, Radi R. HPLC separation of human serum albumin isoforms based on their isoelectric points. J Chromatogr B Analyt Technol Biomed Life Sci. 2014 Jan 1; 944:144-51.
Identificación mediante técnicas inmuno-químicas, mapeos peptídicos; perfiles de N-glicanos y ensayos de Cuantificación mediante métodos colorimétricos y HPLC.
- Manta B, Obal G, Ricciardi A, Pritsch O, Denicola A. Tools to evaluate the conformation of protein products. Biotechnol. J. 2011, Jun; 6(6):731-41.