Recombinant proteins and immunoassays facility
The NGF lab concerns research and development of protocols and methods, is based on recombinant proteins production and immunoassays development, and is involved in different projects carried out at EBRI or in collaboration with other labs.
The NGF lab main activities are:
- Expression and purification of recombinant proteins
NGF lab works on the development and the optimization of protocols for expression and purification of recombinant proteins; in particular, on the production of Nerve Growth Factor (NGF), its precursor proNGF, and their mutant forms with therapeutic prospective such as painless NGF, a candidate for Alzheimer’s Disease therapy and for the treatment of pressure and diabetic ulcers. Furthermore, the NGF Lab produces specific monoclonal antibodies (Mab) such as scFv (single chain variable fragment) anti Aβ oligomers, Mab anti NGF αD11 4GA; MAb anti-trkA MNAC13, MAb anti tau NH2 fragment, that are tools of exclusive property of EBRI. Following the SARS-Cov2 pandemic, NGF Lab is working on the production, purification and biochemical characterization of some scFv anti protein N.
By the means of specific biochemical analyses and bioassays, each batch of purified recombinant protein or antibody is tested in order to verify bioactivity and the batch to batch consistency.
- Immunoassays for measuring biomarkers and therapeutic proteins in biological fluids
NGF lab also develops and validates immunoassays for measuring biomarkers and therapeutic proteins in biological fluids, by using different technologies: ELISA, alphaLISA (Perkin Elmer), Simple WES (Protein Simple). The choice between these different technologies is evaluated case by case, in order to obtain the best assay in terms of sensitivity, reliability, specificity, reproducibility. So far, NGF lab developed and validated i) ELISA to asses pharmacokinetics after drug administration in mouse model and to check antibodies and proteins produced by the facility; iii) alphaLISA to assess pharmacokinetics of NGF painless in human serum and other biological fluids.
NGF Lab has developed a new immunoassay for the measurement of proNGF in human cerebrospinal fluid from Alzheimer’s patients and controls, for the validation of new biomarkers for the early diagnosis of Alzheimer’s disease and other neurodegenerative diseases.
proNGF Measurement in Cerebrospinal Fluid Samples of a Large Cohort of Living Patients With Alzheimer’s Disease by a New Automated Immunoassay.Malerba F, Arisi I, Florio R, Zecca C, Dell’Abate MT, Bruni Ercole B, Camerini S, Casella M, Logroscino G, Cattaneo A. Front Aging Neurosci. 2021 Oct 27;13:741414. doi: 10.3389/fnagi.2021.741414. eCollection 2021. PMID: 34776928. PubMed
The chemokine CXCL12 mediates the anti-amyloidogenic action of painless human nerve growth factor. Capsoni S, Malerba F, Carucci NM, Rizzi C, Criscuolo C, Origlia N, Calvello M, Viegi A, Meli G, Cattaneo A. Brain. 2017 Jan;140(1):201-217. doi: 10.1093/brain/aww271. PubMed
Functional Characterization of Human ProNGF and NGF Mutants: Identification of NGF P61SR100E as a “Painless” Lead Investigational Candidate for Therapeutic Applications. Malerba F, Paoletti F, Bruni Ercole B, Materazzi S, Nassini R, Coppi E, Patacchini R, Capsoni S, Lamba D, Cattaneo A. PLoS One. 2015 Sep 15;10(9):e0136425. doi: 10.1371/journal.pone.0136425. eCollection 2015. PubMed