Here, we report a new manufacturing approach which starts with non-infectious protein and builds structure, and hence antigenicity, without building infectivity. Our approach relies on the expression and purification of viral structural proteins display- ing immunogenic influenza epitopes. These chimeric proteins are prepared in bacteria and then processed in vitro into non-infectious virus-like particles (VLPs) in cell-free downstream process. Protein production can potentially be done in highly efficient bacteria, thus maximising speed and efficiency while also minimising the need for high level biocontainment. This bioprocess potentially provides a rapid response vaccine manufacturing platform for pandemic influenza. References 1. Wilson, J. and von Itzstein, M.: Recent strategies in the search for the new anti- influenza therapies, Curr Drug Targets, 4, (5) 389-408 (Jul 2003). 2. Bardiya, N. and Bae, J.: Influenza vaccines: recent advances in production technologies, Appl Microbiol Biotechnol, 67, (3) 299-305 (2005). doi:10.1016/j.jbiosc.2009.08.099 BP-O2 Freeze-dried crocodile blood production as food supplement Win Chaeychomsri, Sudawan Chaeychomsri, Jindawan Siruntawineti, Duangchan Hengsawadi, and Yaovadee Cuptapun Kasetsart University, Chatuchak Bangkok, Thailand Siamese crocodile is a medicinal animal that has been reported for its blood anti-microbial and antiviral activity. The practice of consuming crocodile blood for improving human health is found in the traditions of many Asian cultures. The large volume-collecting and freeze-drying processes of Siamese crocodile blood were designed and developed. These freeze-dried blood products were packed in capsule, testing for bacterial contamination and heavy metal. The results showed that freeze-dried crocodile blood product have no bacterial and heavy metal contamination. The safety evaluation of freeze-dried crocodile blood diet was per- formed in 5 groups of both sexes of Sprague-Dawley rats. Freeze- dried crocodile blood was orally administered for 24 weeks. The blood ingestion had no effect on rat behavior and survival. After taking the crocodile blood 0, 12, and 24 weeks, the rat blood was collected from tail-vein for hematological investigations. The hematological values did not reveal differences between the treated and the control groups. The biochemical values including alkaline phosphatase (ALP), aspartate transaminase (AST), alanine transaminase (ALT), blood glucose, blood urea nitrogen (BUN), and albumin were not significantly different among experimental groups throughout the study time. Histopathological examinations of the liver intestine and kidney tissue specimens showed no differences in pathological findings between the treated and the control groups. The efficiency of freeze-dried crocodile blood on iron deficiency Sprague-Dawley rat was designed to observe hematological values in rats. The hematological values that indicated anemia status showed the efficiency for promoting hemoglobin and hematocrit values on iron deficiency rat after 4 weeks of treatment and had no detrimental effect on histological change in intestine, kidney, and liver after 24 weeks. These data suggested that the freeze-dried crocodile blood has safety and efficiency for promoting hemoglobin and hematocrit values and may be used as a food supplement in anemia patients. doi:10.1016/j.jbiosc.2009.08.100 BP-O3 Visualization of liposome production in a micro-tube Atsushi Fujiwara, Hiroshi Suzuki, Tomohisa Katsuda, and Yoshiyuki Komoda Department of Chemical Science and Engineering, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe, Japan In the previous study, it was reported that the liposomes produced in a mocro-tube are uniform in size and then the yield becomes very high. The size of liposomes was also reported to be determined with Reynolds number defined by the tube diameter and flow velocity. However, the mechanism of liposome production in such a micro-tube has yet remained to be unknown. In this study, an experimental study on liposome formation in a micro-tube has been performed. Visualization of liposome formation in a micro-tube was conducted with a movable stage set on the microscope system. Figure 1 shows the set-up of experimental apparatus. Ion-exchanged water from a syringe pump flows into a micro-tube where egg yolk lecithin film has pre-formed on the tube wall. The tube size was changed in three steps: 200, 300, and 530 mm and the flow velocity ranged from 50 to 200 μm/s. The microscope stage moved with the flow velocity in order to observe the microscopic formation of liposomes. Figure 2 shows an example of photos at the interface between air and water. At first, the air/water interface peels the lipid film off from the tube surface and a lipid film was re-generated. After this, the attachment of the lipid film onto the tube-wall occurs and the rod-like liposomes are formed as shown in the figure. The diameter of the rod-like liposomes corresponds to the spherical diameter measured outlet of the micro-tube. The rod-like liposomes are enlarged by the water flow in the streamwise direction and divided in the flow by the shear. Thus, the size is concluded to be determined by the Reynolds number. doi:10.1016/j.jbiosc.2009.08.101 BP-O4 High-throughput generation of monoclonal antibodies from single B cells using RT–PCR and a cell-free protein synthesis system Hideo Nakano, and Yunita Sabrina Nagoya University, Nagoya, Japan We have developed a novel method, namely the single-cell RT– PCR-linked in vitro expression (named SICREX), which enables the high-throughput generation and screening of monoclonal antibodies and important biomolecules in immunology and has widespread applications (1). This approach entails the isolation of antibody- producing cells from immunized mouse spleen or human peripheral cells using magnetic microbeads conjugated with B cell- or plasma S22 Abstracts / Journal of Bioscience and Bioengineering 108 (2009) S21–S28