Biotechnology encompasses everything from the production of recombinant proteins to the use of biological molecules as components of nano-technology. Fundamental studies in various aspects of biology can lead to a variety of possible applications.

Development of therapeutic pharmaceuticals
Proteins participate in homeostasis through the interaction of a specific part of the surface with other proteins, nucleic acids, carbohydrates and other ligands. Once these interacting sites have been mapped and their function understood, agonists or antagonists can be designed and developed for use as therapeutic agents. The physiologically important functional site can be grafted on a small protein scaffold using protein engineering to enhance the stability and to retain specific conformation. Our department has several undergoing programs which are attempting to develop therapeutic prototypes based on the structure-function relationships of biologically important proteins; for example the group led by A/P Manjunatha Kini is developing therapeutic agents based on toxins isolated from snake venoms. Simple invertebrates and vertebrates offer invaluable hints in the development of drugs for human healthcare. One example is the use of antimicrobial properties of recombinant Factor C from the horseshoe crab, which is currently being commercialized by Cambrex as diagnostics for endotoxin, following the research by Prof Ding Jeak Ling and her group. The same group has developed the use of Factor C-derived Sushi peptides for high performance chromatographic separation and purification, for the removal of endotoxin from biological fluids and biomedical therapeutics for human parenteral administration: the patent for this has been filed and has been allowed in principle. The same group has also developed a recombinant yeast-vitellogenin as SCP (Single Cell protein), as novel live-feed for larvae, and the patent has been filed. Dr Chew Fook Tim's laboratory has been able to identify and produce recombinant proteins to the full spectra of allergens from most of the major species of house dust mites, the main source of allergenic components worldwide. These have now been used in several studies worldwide for component resolved diagnosis of allergies and are being developed further as hypoallergenic vaccines for immunotherapeutic uses to alleviate such disease conditions. A number of groups have also been working on the SARS virus in order to develop early diagnostic tools, for example in the laboratory of A/P Wong Sek Man through use of monoclonal antibody technology.

Plant cells and plant viral vectors
As a natural genetic engineer, Agrobacterium tumefaciens can deliver T-DNA into different eukaryotes, including plant, yeast, fungal and human cells. The gene transfer is facilitated by a pilot protein VirD2 that guides the transfer of T-DNA inside both bacterial and eukaryotic cells. A/P Pan Shen Quan's group has identified a eukaryotic protein that interacts with VirD2 and is involved in the trafficking of the T-DNA inside eukaryotic cytoplasm. They have also developed a method that can detect single T-DNA molecules inside eukaryotic cells, presenting them with a unique and effective position to illustrate the T-DNA trafficking pathway inside the eukaryotic cells. Based on their knowledge on the transfer process, they are currently designing and developing novel Agrobacterium-based DNA delivery systems for gene therapy, and hope to develop novel gene and protein delivery systems for various organisms. The same group has also developed an Agrobacterium cell surface display system that may be used to display different proteins on bacterial surface. Our system may display large and complex proteins with quaternary structure and disulfide bonds. The display may be regulated by a simple parameter such as acidic pH. Based on these unique features, they are developing cell-based vaccines and engineer proteins of various applications in biotechnology.

Dr. Yu Hao's group has conducted the successful genetic transformation of a Dendrobium orchid via Agrobacterium tumefaciens, which was the first case in the world showing successful transfer of a complete target gene into orchids using Agrobacterium. By using this system, they are now investigating a series of developmental genes involved in orchid flower development. These studies will improve the genetic manipulation of gene activities to change important traits in orchids. Plant viral vectors are also being developed by the group of A/P Wong Sek Man, in order to express useful peptides or proteins related to biomedical research. The same group is working to identify novel internal ribosome binding sequences for improvement of gene expression systems and to generate virus-resistant orchids and watermelons. A strategic project on using plant cells as bioreactors for production of biomedically relevant proteins has also been initiated recently by the group of A/P Kumar. These proteins generally require elaborate post-translational modifications specific for eukaryotes. Hence, the recombinant proteins produced using bacterial systems tend to lack desired biological functions. The current work focuses on selected genes that have potential to be used as diagnostics for detecting allergens (e.g., from the dust mite) or have therapeutic potential (sequences based on snake venom proteins).

Biosensors, biomonitors and transgenics
The fluorescent transgenic zebrafish, produced by the group of A/P Gong Zhiyuan, are being marketed in USA with the trade name GloFishTM, which are widely praised as the first public accessible transgenic pet and have been increasingly used asan educational model for transgenic technology. Currently, the same group is employing the transgenic technology in two small model aquarium fish, the zebrafish ( Danio rerio) and medaka ( Oryzias latipes), for several major applications including generation of fluorescent ornamental fish, biomonitoring fish, bioreactor fish and oncofish. They are also developing zebrafish DNA chip to be used as a biomonitoring tool for detection of environmental pollutants and understanding molecular basis of toxicity.

Bio-nanotechnolgy research
'Nano' is the unit of size that refers to the scale of one billionth of one meter. This is the diameter of a few ten folds of atom and is the size of individual, important biological molecules, such as proteins, nucleic acids, lipids, sugars that are the building blocks of living organisms. Scientists are beginning to discover and understand new properties of materials that have been realized for preparation and manipulation at the size of nanometer. Hence, we are in a new era of fulfilling the predictive statement in 1959 that 'there are plenty rooms at the bottom' by Dr. Richard Feynman, 1965 Nobel Laureate in Physics, as in that “the principles of physics, as far as I can see, do not speak against the possibility of maneuvering things atom by atom.' The electrochemical biosensor laboratory led by A/P Sheu Fwu-Shan, group together disciplines in Biological Sciences, Chemical and Physical Sciences to devote themselves in bio-nanoscience research by exploring and studying the properties of the nano-scale materials. Furthermore, they wish to translate the unique properties of nano-scale materials to technology by preparing materials, devicesand systems through controlling of matter on the length scale of approximately 1-100 nanometer range. Ultimately, there will be exploitation of novel properties and phenomena developed at that scale for useful applications. The most promising applications they had so far found are the establishment of some biosensor devices made by nano-scale matters such as carbon nanotubes and nano metal particle-lipid membrane composite suitable for fast and reliable detection of biological molecules including nitric oxide, oxygen, glucose, uric acid, ascorbic acid and neurotransmitter dopamine. The group of A/P Lim Tit Meng and colleagues from the Paediatrics Department and the Institute of Microelectronics have developed a series of micro-PCR devices based on micro-electro-mechanical systems (MEMS) technology. The device meets the requirements for reproducibility in the temperature ramping and cooling cycles, compatibility with the PCR reactions, and the robustness in amplifying gene products of genomic source. In fact the micro-PCR has many features that are superior to the conventional PCR as well as some devices of the same miniaturized nature. After the successful development of a single chamber micro-PCR on chip, they went on to develop a multiplex thermal cycler fabricated in a micro-assembly manner using flip-chip bonding technique, which is batch manufacturable with good reproducibility. At least three USA patents have been granted for the inventions. The group aims to develop labon-chip devices integrated for DNA/RNA sample preparation to amplification to detection using BioMEMS
and nanotechnology.