Immobilization Of Hybridoma Cells In Sol-GEL Silica Matrices.

DESIMONE M; M. C. DE MARZI; G. ALVAREZ; E. L. MALCHIODI; L. E. DIAZ

Francia

Conferencia; First International Conference on Multifunctional, Hybrid and Nanomaterials; 2009

The encapsulation of biosystems, including proteins, antibodies, enzymes, bacteria, and mammalian whole cells is becoming popular in several fields such as drug delivery, immunoaffinity chromatography, building of biosensors or bioreactors, and cell encapsulation for bioartificial organs or tissue engineering. At the forefront of research in this area is the development of methods for treatment, diagnosis, monitoring and control of biological systems. The sol-gel process is an inorganic polymerization taking place, in some cases, in mild conditions allowing the association of mineral phases with organic or biological systems. Optimization of the technological parameters affecting the mechanical properties and permeability of sol-gel matrices is essential to develop matrices with improved properties for cell immobilization. In this work, the effect of different parameters such as the concentration of different sol-gel precursors, including alkoxide, aqueous and polyol-modified silanes routes, and immobilized cell density, on the function of hybridoma cells producing anti-peptidoglycan receptor protein (PGRP) I-á. monoclonal antibody (mAb) has been investigated. Cell morphology was conserved as it was observed by optical and fluorescence microscopy. Cellular activity of immobilized hybridoma cells was determined by the tetrazolium assay (MTT assay) and a LIVE/DEAD cell vitality fluorescent assay. Viability was 89.5% in tetrakis (2-hydroxyethyl) ortosilitate-derived matrices and was similar to the values obtained in sodium silicate derived matrices. Significant differences were observed on cellular activity between the various cells densities employed in this work (2,5x105 – 1,5x106 cells/ml), being higher for lower cell densities when expressed on a per cell basis. MAb production from immobilized cells, was detected in high levels by ELISA even after 15 days post immobilization. The design of such novel devices with significant added value is a key factor when foreseeing industrial developments of sol-gel materials in medicinal science