Modulation of hybridoma cell growth and antibody production by coating cell culture material with extracellular matrix proteins - Monoclonal Antibodies

Modulation of hybridoma cell growth and antibody production by coating cell culture material with extracellular matrix proteins
Received 18 May 2006; revised 21 December 2006; accepted 16 January 2007. Available online 11 February 2007
K. Heilmanna, T. Grothb, c, M. Schossigc, A. Lendleinc and B. Micheel a
Biochemical Engineering Journal
Volume 35, Issue 3, 1 August 2007
ScienceDirect
Copyright ? 2007 Elsevier B.V. All rights reserved.
aDepartment of Biotechnology, Institute of Biochemistry and Biology, University of Potsdam, Karl-Liebknecht-Strasse 24-25, D-14476 Golm, Germany
bBiomedical Materials Group, Institute of Pharmaceutical Technology and Biopharmacy, University of Halle-Wittenberg, D-06099 Halle (Saale), Germany
cGKSS Research Centre, Institute of Polymer Research, Kantstrasse 55, D-14513 Teltow, Germany
Abstract
The influence of coating polystyrene tissue culture plates with different proteins on murine hybridoma cell growth and antibody production was investigated. Fibronectin, collagen I, bovine serum albumin and laminin were used to coat NUNC? and COSTAR? cell culture plates. Cell number and antibody concentration in culture fluids were quantified as indicators for cell viability, proliferation and productivity. Adhesive behaviour, morphology, expression of surface receptors of hybridoma cells and the presence of tyrosine-phosphorylated proteins in cell lysates were characterized by cell adhesion experiments, microscopy, flow cytometry and Western Blot analysis.
It was shown that coatings with fibronectin (0.2 ?g/ml) lead to a substantial improvement of cell growth by 50?70% and an increase of monoclonal antibody production by 100?120%.
Collagen I coatings showed an improvement in cell growth by 30?70% and by 60% for the production of monoclonal antibodies. Coatings with BSA and laminin had minor effects on these parameters. It was found that the hybridoma cell lines used in this study did not express the a2-chain of the a2?1-integrin, which is responsible for binding to collagen and laminin.
However, the presence of ?1-integrin on the cell surface was shown, which should enable hybridoma cells to bind fibronectin. We propose, therefore, that fibronectin adsorption to cell culture materials may be a promising approach to enhance the production of monoclonal antibodies by cultivated hybridoma cells.
Keywords: Hybridoma cells; Tissue culture plates; Cell growth; Antibody production; Extracellular matrix; Fibronectin adsorption
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