Expression and production of llama variable heavy-chain antibody fragments (VHHs) by Aspergillus awamori
作者： Vivi Joosten , Robin J. Gouka , Cees A. M. J. J. van den Hondel , C. Theo Verrips , B. Christien Lokman
DOI: 10.1007/S00253-004-1689-0
关键词:
摘要: We report the expression and production of llama variable heavy-chain antibody fragments (VHHs) by Aspergillus awamori. Fragments encoding VHHs were cloned in a suitable vector transformants secreting VHH analysed for integrated gene copy-numbers, mRNA levels protein production. Functional detected culture medium, indicating feasibility producing this type fungal system. Secreted subjected to (extracellular) degradation, which could be partially prevented addition BSA medium.
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Peter J. Punt, Cees A.M.J.J. van den Hondel, Transformation of filamentous fungi based on hygromycin B and phleomycin resistance markers. Methods in Enzymology. ,vol. 216, pp. 447- 457 ,(1992) , 10.1016/0076-6879(92)16041-H
Elvin A. Kabat, Sequences of proteins of immunological interest National Institutes of Health. ,(1991)
Robin J. Gouka, Casper Gerk, Paul J.J. Hooykaas, Paul Bundock, Wouter Musters, C. Theo Verrips, Marcel J.A. de Groot, Transformation of Aspergillus awamori by Agrobacterium tumefaciens-mediated homologous recombination. Nature Biotechnology. ,vol. 17, pp. 598- 601 ,(1999) , 10.1038/9915
Margaret Alic, J. W. Bennett, Linda L. Lasure, More gene manipulations in fungi Academic Press. ,(1991)
M J Hynes, C M Corrick, J A King, Isolation of genomic clones containing the amdS gene of Aspergillus nidulans and their use in the analysis of structural and regulatory mutations. Molecular and Cellular Biology. ,vol. 3, pp. 1430- 1439 ,(1983) , 10.1128/MCB.3.8.1430
Haruyo Sawai-Hatanaka, Toshihiko Ashikari, Yoshikazu Tanaka, Yasuhiko Asada, Toru Nakayama, Hiroyuki Minakata, Naoki Kunishima, Keiichi Fukuyama, Hideaki Yamada, Yuji Shibano, Teruo Amachi, Cloning, Sequencing, and Heterologous Expression of a Gene Coding for Arthromyces ramosus Peroxidase Bioscience, Biotechnology, and Biochemistry. ,vol. 59, pp. 1221- 1228 ,(1995) , 10.1271/BBB.59.1221
Margareta Kolar, Peter J. Punt, Cees A.M.J.J. van den Hondel, Helmut Schwab, Transformation of Penicillium chrysogenum using dominant selection markers and expression of an Escherichia coli lacZ fusion gene. Gene. ,vol. 62, pp. 127- 134 ,(1988) , 10.1016/0378-1119(88)90586-0
R. J. Gouka, J. G. M. Hessing, H. Stam, W. Musters, C. A. M. J. J. van den Hondel, A novel strategy for the isolation of defined pyrG mutants and the development of a site-specific integration system for Aspergillus awamori. Current Genetics. ,vol. 27, pp. 536- 540 ,(1995) , 10.1007/BF00314444
R.H.J. van der Linden, L.G.J. Frenken, B. de Geus, M.M. Harmsen, R.C. Ruuls, W. Stok, L. de Ron, S. Wilson, P. Davis, C.T. Verrips, Comparison of physical chemical properties of llama VHH antibody fragments and mouse monoclonal antibodies. Biochimica et Biophysica Acta. ,vol. 1431, pp. 37- 46 ,(1999) , 10.1016/S0167-4838(99)00030-8
M.S. Blake, K.H. Johnston, G.J. Russell-Jones, E.C. Gotschlich, A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots Analytical Biochemistry. ,vol. 136, pp. 175- 179 ,(1984) , 10.1016/0003-2697(84)90320-8