Modulation of hybridoma cell growth and antibody production by coating cell culture material with extracellular matrix proteins |
| |
| Institution: | 1. Department of Biotechnology, Institute of Biochemistry and Biology, University of Potsdam, Karl-Liebknecht-Strasse 24-25, D-14476 Golm, Germany;2. Biomedical Materials Group, Institute of Pharmaceutical Technology and Biopharmacy, University of Halle-Wittenberg, D-06099 Halle (Saale), Germany;3. GKSS Research Centre, Institute of Polymer Research, Kantstrasse 55, D-14513 Teltow, Germany;1. School of Medicine, Nankai University, 94 Weijin Road, Tianjin 300071, China;2. Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Huanhu West Road, Tianjin 300060, China;1. Dokuz Eylul University, Faculty of Medicine, Department of Medical Biology, Inciralti, 35340 Izmir, Turkey;2. Dokuz Eylul University, Izmir Biomedicine and Genome Center, Inciralti, 35340 Izmir, Turkey;3. Dokuz Eylul University, Faculty of Medicine, Department of Pharmacology, Inciralti, 35340 Izmir, Turkey;1. Department of Pharmacology, Medical University of Silesia, Medyków 18, 40-752 Katowice, Poland;2. Faculty of Chemistry, University of Gdańsk, Wita Stwosza 63, 80-952 Gdańsk, Poland;3. Department of Neurology, Henry Ford Hospital, 2799 West Grand Boulevard, Detroit, MI 48202, USA;1. Department of Biomedical Sciences, City University of Hong Kong, Tat Chee Avenue, Kowloon Tong, Hong Kong;2. Institute of Bioengineering, Zhejiang University of Technology, 18 Chaowang Road, Hangzhou 310014, China |
| |
| 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 μg/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 α2-chain of the α2β1-integrin, which is responsible for binding to collagen and laminin.However, the presence of β1-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: | |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
