Recent Patents on Biotechnology (v.6, #1)

Preface by Rongling Wu (1-1).
As a cutting-edge, peer-reviewed journal launched by Bentham Science, Recent Patents on Biotechnology (BIOT) covers the science and engineering of biotechnology. It publishes new advances in technology and methodology of relevance to the biological, biomedical, agricultural and environmental sciences. It also provides researchers with special issues on timely topics in the field of biotechnology. The journal includes articles ranging from the review to research, and it covers all levels of biological organization. Representative topics of interest include: molecular engineering of nucleic acids and proteins; molecular therapy using therapeutics genes, antisense, siRNAs, aptamers, DNAzymes, ribozymes, peptides, and proteins; systems biology related with genomics, functional genomics, proteomics, structural genomics, metabolomics, etc.; algorithms and modeling in computational biology; regenerative medicine; imaging informatics; food and agricultural biotechnology; and environmental biotechnology. The journal provides a focus for communication among biotechnology researchers who study biotechnologies and use them as a tool to understand basic sciences or solve practical problems in biomedicine, agriculture and environmental protection. The journal is an international forum for the exchange of novel and significant information gained from empirical and theoretical investigations on the innovation and application of new biotechnologies. BIOT seeks contributions from scholars and scientists from around the world. BIOT also welcomes leading experts and experienced researchers to sponsor special issues in their areas. As the Editor-in-Chief of BIOT, I wish to work closely with our editorial advisory board and authors to establish the scientific reputation of the journal and elevate and maintain its international impact in the years to come. Lastly, but not least, I wish to thank Bentham Science Publishers for providing me with such a platform to serve our biotechnology community and Ms. Sadia Haris and Ms. Donia Akhter for their tireless support on my editorial work.

Numerous bacterial functions, such as virulence and biofilm formation, are controlled by a cell densitydependent communication mechanism known as Quorum Sensing (QS), in which small diffusible molecules are released, allowing bacteria to coordinate their behavior once a minimal effective quorum has been reached. The interference with these signaling systems, also known as Quorum Quenching (QQ), represents a promising strategy to tackle bacterial infections. The growing interest in this approach is reflected by the increasing number of patents within the field (45 up to now), especially in the last few years, as shown by patent applications published since 2009. The fact that biofilm formation is also controlled by QS systems expands the application of QQ to clinically-relevant biofilms such as those responsible for periodontal disease. Moreover, since biofilms increase bacterial resistance to antimicrobials, QQ could represent a new way to fight some of the most recurrent human pathogens, such as nosocomial multiresistant strains, and this deserves further exploration, especially through more proofs of concept. In this article we review the best known QS and QQ systems to date and we describe recent patents on the interference with this type of bacterial communication.

Biofilms: New Ideas for An Old Problem by Carla C.C.R. de Carvalho (13-22).
Microbial cells, under moist conditions, are able to adhere to surfaces and to form structured communities embedded in a matrix of extracellular polymeric substances (EPS). In industrial environments, biofilms can cause heat and mass transfer limitations whilst in medical facilities they can be a source of contamination and proliferation of infections. Biofilm formation is related to the pathogenicity of some bacterial strains and cells in biofilms are usually resistant to antimicrobials agents, which increases the interest in new and sound methods for their prevention and destruction.

The expression in Escherichia coli strain Rosetta of the recombinant acidic subunit from the 11S amaranth seed storage protein (protein ACM3) was studied at flask and at bioreactor levels. This subunit was modified by inserting four Val-Tyr antihypertensive peptides in tandem into its third variable region and also with the tripeptide Ile-Pro-Pro in the Cterminal region. Flasks experiments allowed us to define the best conditions for the preparation and expression and accumulation of the protein ACM3, including the certainty of its presence within the cells especially as an insoluble fraction. The effects of cultivation temperature, aeration rate and agitation speed on the production of the protein ACM3 was tested in a 5-L batch bioreactor. Applying response surface methodology (RSM) we found that the aeration rate was the most significant factor affecting in a positively way the production yields and productivity of the recombinant protein. Temperature had effect only in conjunction to aeration. The highest recombinant acidic subunit concentration (747 mg L-1) and the highest productivity (186 mg L-1 h-1) were attained in 4 h of cultivation when the factors evaluated were controlled at its central values: 0.1 vvm, 300 rpm, and 30.5° C. Results from this study indicate that RSM is an effective technique to maximize the production of this recombinant protein.

Laboratory Dialysis-Past, Present and Future by Parikshit Bansal (32-44).
Laboratory dialysis, one of the most widely used techniques in biological research is truly a ‘ gateway technology’ . The analogy is to that of a ‘ gate’ of a building through which everybody has to pass, even though they may wish to go to different departments. Similarly, researchers may be working in altogether different areas but all may need to use laboratory dialysis at one stage or the other during the course of their research. Biochemists may use it to purify enzymes, an immunologist may use it to purify monoclonal antibodies from culture supernatants, a chemist may use it as a step in the crystallography process or for purification of ionic liquids, a biotechnologist may use it to study the effectiveness of enzyme immobilization and a drug discovery scientist may use it for determining drug-protein interaction. The present article reviews patents in the field of laboratory dialysis from inception till date, focusing on the various developmental and innovation related milestones during evolution of the technique. It captures the full panorama of a very interesting technique which continues to be as relevant today as it was in 1866 when the term ‘ dialysis’ was first coined.

Three main targets were subjected for the most approved monoclonal antibodies (mAbs) in cancer therapy: EGFR in solid cancer, the clusters of differentiation in blood cancer and VEGF in angiogenesis. Meanwhile side effects, the elevated costs and resistance problems are limiting the efficiency of mAbs as targeted therapy. The combinatory therapy with chemo or radiotherapy has improved the efficiency of mAbs. The present review aims to shed more light on the immunotherapy and the related patents that were developed for cancer treatment.

Overexpression of recombinant proteins in bacterial systems (such as E. coli) often leads to formation of inactive and insoluble ‘ inclusion bodies’ . Protein refolding refers to folding back the proteins after solubilizing/unfolding the misfolded proteins of the inclusion bodies. Protein aggregation, a concentration dependent phenomenon, competes with refolding pathway. The refolding strategies largely aim at reducing aggregation and/or promoting correct folding. This review focuses on non-chromatographic strategies for refolding like dilution, precipitation, three phase partitioning and macro-(affinity ligand) facilitated three phase partitioning. The nanomaterials which disperse well in aqueous buffers are also discussed in the context of facilitating protein refolding. Apart from general results with these methods, the review also covers the use of non-chromatographic methods in protein refolding in the patented literature beyond 2000. The patented literature generally describes use of cocktail of additives which results in increase in refolding yield. Such additives include low concentration of chaotropic agents, redox systems, ions like SO4 2- and Cl-, amines, carboxylic acids and surfactants. Some novel approaches like use of a “pressure window” or ionic liquids for refolding and immobilized diselenide compounds for ensuring correct

During the past epoch we have gone through the remarkable progress in plant gene transformation technology. The production of transgenic plants is considered as a valuable tool in plant research and the technology is extensively applied in phytomedicines and agricultural research. Gene transformation in plants is normally carried out by Agrobacterium species, application of some chemicals and physical techniques (electroporation, microprojectile, etc.). Now a days with better efficacy and reproducibility, novel technologies for the direct gene transfer like liposome, positively charged liposome (lipofectin) and nanoparticle based delivery systems are used for genetic transformation of plants. In this review, we have enlightened the novel nanotechnologies like liposome, Carbon nano-tube and nanoparticles with their current status and future prospects in transgenic plant development. Moreover, we have also highlighted the limitations of conventional techniques of gene transfer. Furthermore, we have tried to postulate innovative ideas on the footprints of established nanotechnology and chemical based strategy with improved efficacy, reproducibility and accuracy along with less time consumption.

Patent Selections by Mehrnaz S. Ohadi Rafsanjani (80-80).
The patents annotated in this section have been selected from various patent data bases. These recent patents are relevant to the articles published in this journal issue, categorized by different biotechnology methods, processes and techniques involved.