Recent Patents on Biotechnology (v.5, #3)

Recent progress in the development of anthrax vaccines by Manpreet Kaur, Rakesh Bhatnagar (148-159).
Bacillus anthracis is the etiological agent of anthrax. Although anthrax is primarily an epizootic disease; humansare at risk for contracting anthrax. The potential use of B. anthracis spores as biowarfare agent has led to immenseattention. Prolonged vaccination schedule of current anthrax vaccine and variable protection conferred; often leading tofailure of therapy. This highlights the need for alternative anthrax countermeasures. A number of approaches are being investigatedto substitute or supplement the existing anthrax vaccines. These relied on expression of Protective antigen(PA), the key protective immunogen; in bacterial or plant systems; or utilization of attenuated strains of B. anthracis forimmunization. Few studies have established potential of domain IV of PA for immunization. Other targets including thespore, capsule, S-layer and anthrax toxin components have been investigated for imparting protective immunity. It hasbeen shown that co-immunization of PA with domain I of lethal factor that binds PA resulted in higher antibody responses.Of the epitope based vaccines, the loop neutralizing determinant, in particular; elicited robust neutralizing antibodyresponse and conferred 97% protection upon challenge. DNA vaccination resulted in varying degree of protectionand seems a promising approach. Additionally, the applicability of monoclonal and therapeutic antibodies in the treatmentof anthrax has also been demonstrated. The recent progress in the direction of anthrax prophylaxis has been evaluated inthis review.

Miniaturization in Biotechnology: Speeding up the Development of Bioprocesses by Pedro Fernandes, Filipe Carvalho, Marco P.C. Marques (160-173).
The use of miniaturized devices for fastening bioprocess development, even up to production scale, has expandedrapidly, a feature clearly noticeable in recent years. This matter was reviewed in a recent past, but several developmentshave occurred since. These will be addressed in the present work, which will provide some insight on the use ofmicrofluidic /microstructured reactors and of micro-scale downstream processing as well, therefore broadening the scopeof the review.

The Offer of Chemistry to Targeted Therapy in Cancer by Ikram Jemel, Karim Jellali, Jihene Elloumi, Sami Aifa (174-182).
Cancer therapy is facing the big challenge of destroying selectively tumour cells without harming the normaltissues. Chemotherapy was trying from the beginning to kill malignant cells because of their proliferative activity sincenormal cells are in general quiescent. Meanwhile side effects were produced due to the destruction of some normal cellsthat need regular proliferation. The discovery of biomarkers led to the identification of molecular targets within tumourcells in order to kill them selectively. Chemistry followed the progress of biomarkers biotechnology by the production oftarget specific antagonists which were the subject of many patents. Meanwhile novel problems of tumour resistance appearedand made the battle against cancer a non stop development of new strategies and new weapons. As a consequence,paralleled activities of patenting biomarkers and chemical antagonists are continuously generated. The offer of chemistrydoes not actually limit the efficiency of Targeted therapy but the identification of biomarkers is still missing the exclusivespecificity to tumour cells.

Recent Achievements on Siderophore Production and Application by Carla C.C.R. de Carvalho, Marco P.C. Marques, Pedro Fernandes (183-198).
Iron is the most abundant chemical element on Earth but its most common oxidation state is Fe(III) which presentsa very low solubility under physiological conditions. During evolution, micro-organisms have developed soundstrategies to acquire iron from both the environment and superior organisms, including direct uptake of iron ions from exogenousiron/heme sources and the synthesis of specialized Fe(III) chelators called siderophores. The present review paperaims at presenting and discussing the latest achievements in siderophore isolation and production, as well as novel applicationsof these molecules in therapies against iron-related diseases and in vaccines, and their application as antimicrobialagents and biosensors.

Native and Heterologous Production of Bacteriocins from Gram-Positive Microorganisms by Mabel Munoz, Diana Jaramillo, Adelina del Pilar Melendez, Carlos J. Almeciga-Diaz, Oscar F. Sanchez (199-211).
In nature, microorganisms can present several mechanisms for setting intercommunication and defense. One ofthese mechanisms is related to the production of bacteriocins, which are peptides with antimicrobial activity. Bacteriocinscan be found in Gram-positive and Gram-negative bacteria. Nevertheless, bacteriocins produced by Gram-positive bacteriaare of particular interest due to the industrial use of several strains that belong to this group, especially lactic acid bacteria(LAB), which have the status of generally recognized as safe (GRAS) microorganisms. In this work, we will reviewrecent tendencies in the field of invention and state of art related to bacteriocin production by Gram-positive microorganism.Hundred-eight patents related to Gram-positive bacteriocin producers have been disclosed since 1965, from which57% are related bacteriocins derived from Lactococcus, Lactobacillus, Streptococcus, and Pediococcus strains. Surprisingly,patents regarding heterologous bacteriocins production were mainly presented just in the last decade. Although themajor application of bacteriocins is concerned to food industry to control spoilage and foodborne bacteria, during the lastyears bacteriocin applications have been displacing to the diagnosis and treatment of cancer, and plant disease resistanceand growth promotion.

Recent Patents on Alphavirus Protein Expression and Vector Production by Alejandro Aranda, Marta Ruiz-Guillen, Jose I. Quetglas, Jaione Bezunartea, Erkuden Casales, Cristian Smerdou (212-226).
Alphaviruses contain a single-strand RNA genome that can be modified to express heterologous genes at highlevels. Alphavirus vectors can be packaged within viral particles (VPs) or used as DNA/RNA layered systems. The broadtropism and high expression levels of alphavirus vectors have made them very attractive for applications like recombinantprotein expression, vaccination or gene therapy. Expression mediated by alphavirus vectors is generally transient due toinduction of apoptosis. However, during the last years several non-cytopathic mutations have been identified within thereplicase sequence of different alphaviruses, allowing prolonged protein expression in culture cells. Some of these mutants,which have been patented, have allowed the generation of stable cell lines able to express recombinant proteins forextended periods of time in a constitutive or inducible manner. Production of alphavirus VPs usually requires cotransfectionof cells with vector and helper RNAs providing viral structural proteins in trans. During this process full-length wildtype (wt) genomes can be generated through recombination between different RNAs. Several new strategies to reduce wtvirus generation during packaging, optimize VP production, increase packaging capacity, and provide VPs with specifictargeting have been recently patented. Finally, hybrid vectors between alphavirus and other types of viruses have led to anumber of patents with applications in vaccination, cancer therapy or retrovirus production.

In vitro Propagation of Cichorium intybus L. and Quantification of Enhanced Secondary Metabolite (Esculin) by Mehrnaz S. Ohadi Rafsanjani, Amene Alvari, Anis Mohammad, M.Z. Abdin, M.A. Hejazi (227-234).
In this report, rapid and effective shoot as well as root regeneration system through direct multiplication wassuccessfully developed for Cichorium intybus L. Furthermore, the effect of exogenous growth regulators (TDZ and IAA)at different concentrations on the regulation process of the plant was also studied. Enhanced production of esculin in developedC. intybus L. was evaluated using leaf extract. Only on the expense of 20 days, regeneration was seen and verylow dose of TDZ was seen to be more effective. When 0.02mg/L of TDZ was combined with 1.5mg/L of IAA, nearly100% of explants produced shoots with the highest number of regenerated shoots (85.37). With further increase in concentration(..0.05mg/L), the number of shoots per explants get decreased. A lower NAA to IBA ratio (1.0mg/L of IBA and0.5mg/L of NAA) seemed to be more effective for root generation and considered to be the most effective combinationamong the tried groups. IBA was more effective in root development than NAA, but both were comparatively effective.On quantitative analysis by RP-HPLC, the 76.23% of Esculin were found in leaf extract of the in vitro developed C. intybusL. This amount was 26.77% higher than normal grown plants.

Patent Selections by Bentham Science Publishers (235-236).
The patents annotated in this section have been selected from various patent data bases. These recent patents are relevant tothe articles published in this journal issue, categorized by different biotechnology methods, processes and techniques involved.