Direct kinetic assay of interactions between small peptides and immobilized antibodies using a surface plasmon resonance biosensor

• Type: Article in International Scientific Journal
• Date: 2002

Authors

Gomes, P (Author) FCUP; Andreu, D (Author) Other

Journal

Title: Journal of Immunological Methods

Vol. 259

Pages: 217-230

ISSN: 0022-1759

Publisher: Elsevier

Indexing

ISI Web of Knowledge - 36 Citations

Scopus - 40 Citations

Scientific classification

FOS: Medical and Health sciences > Basic medicine

Other information

Authenticus ID: P-000-RF0

DOI: 10.1016/s0022-1759(01)00503-8

Abstract (EN): A surface plasmon resonance (SPR) protocol is described for the direct kinetic analysis of small antigenic peptides interacting with immobilized monoclonal antibodies (mAb). High peptide concentrations (up to 2.5 muM) and medium mAb surface densities (about 1.5 ng/mm(2)) are needed to ensure measurable binding levels, and fast buffer flow rates (60 mul/min) are required to minimize diffusion-controlled kinetics. Good reproducibility levels in the kinetic constants are obtained under these analysis conditions (standard deviations below 10% of the mean values). Application of this protocol to determine the antigenic ranking of viral peptides shows an excellent agreement between SPR and previous competition enzyme-link-ed immunosorbent assays (ELISA) on the same peptide/antibody systems.

Language: English

Type (Professor's evaluation): Scientific