Recent Patents on Biotechnology (v.7, #1)

Preface by Rongling Wu (1-1).

A Review of Recent Patents on the Protozoan Parasite HSP90 as a Drug Target by Sergio O. Angel, Mariana Matrajt, Pablo C. Echeverria (2-8).
Diseases caused by protozoan parasites are still an important health problem. These parasites can cause a widespectrum of diseases, some of which are severe and have high morbidity or mortality if untreated. Since they are still uncontrolled,it is important to find novel drug targets and develop new therapies to decrease their remarkable social andeconomic impact on human societies. In the past years, human HSP90 has become an interesting drug target that has ledto a large number of investigations both at state organizations and pharmaceutical companies, followed by clinical trials.The finding that HSP90 has important biological roles in some protozoan parasites like Plasmodium spp, Toxoplasmagondii and trypanosomatids has allowed the expansion of the results obtained in human cancer to these infections. Thisreview summarizes the latest important findings showing protozoan HSP90 as a drug target and presents three patents targetingT. gondii, P. falciparum and trypanosomatids HSP90.

Although many patents reported bioactive peptides with numerous demonstrated bioactivities and potentialapplications, there exist some limitations to the production of large quantities to satisfy the growing market demands.Indeed, considering that most functional peptides are present in complex matrices containing a large number of hydrolyzedprotein fractions, their separation and purification are required. Some advances have been made in the use of conventionalpressure-driven processes for the continuous production and separation of peptides, however, most of these patentedtechnologies are not scalable and demonstrate a low selectivity when separating similar sized biomolecules. To improvethe separation efficiency, the use of an external electric field during pressure-driven filtration was proposed and patented.However, whatever the claims, the pressure gradient brings about the accumulation of peptides at the nearby membranesurface and affects the membrane transport selectivity. To overcome these drawbacks, a recent patent proposed thesimultaneous fractionation of acidic and basic peptides, using a conventional electrodialysis cell, in which some ion exchangemembranes are replaced by ultrafiltration ones. The perspectives in the field of peptide separation will be the developmentof new membrane materials and new equipments such as microfluidic devices to improve selectivity and yieldof production.

Stabilization and Delivery Approaches for Protein and Peptide Pharmaceuticals: An Extensive Review of Patents by Suryakanta Swain, Debanik Mondal, Sarwar Beg, Chinam Niranjan Patra, Subas Chandra Dinda, Jammula Sruti, Muddana Eswara Bhanoji Rao (28-46).
Proteins and peptides are the building blocks of human body and act as the arsenal to combat against the invadingpathogenic organisms for treatment and management of diseases. Majority of such biomacromolecules are synthesizedby the human body itself. However, entry of disease causing pathogens causes misleading in the synthesis of desired proteinsfor antibody formation. In such alarming situations, the delivery of requisite protein and peptide from external sourcehelps in augmenting the body

Although thermal sterilisation is a widely employed industrial process, little work is reported in the availableliterature including patents on the mathematical analysis and simulation of these processes. In the present work, softwarepackages have been developed for computer aided optimum design of thermal sterilisation processes. Systems involvingsteam sparging, jacketed heating/cooling, helical coils submerged in agitated vessels and systems that employ externalheat exchangers (double pipe, shell and tube and plate exchangers) have been considered. Both batch and continuous operationshave been analysed and simulated. The dependence of del factor on system / operating parameters such as massor volume of substrate to be sterilised per batch, speed of agitation, helix diameter, substrate to steam ratio, rate of substratecirculation through heat exchanger and that through holding tube have been analysed separately for each mode ofsterilisation. Axial dispersion in the holding tube has also been adequately accounted for through an appropriately definedaxial dispersion coefficient. The effect of exchanger characteristics/specifications on the system performance has alsobeen analysed. The multiparameter computer aided design (CAD) software packages prepared are thus highly versatile innature and they permit to make the most optimum choice of operating variables for the processes selected. The computedresults have been compared with extensive data collected from a number of industries (distilleries, food processing andpharmaceutical industries) and pilot plants and satisfactory agreement has been observed between the two, thereby ascertainingthe accuracy of the CAD softwares developed. No simplifying assumptions have been made during the analysisand the design of associated heating / cooling equipment has been performed utilising the most updated design correlationsand computer softwares.

Overexpression of a Modified Amaranth Protein in Escherichia coli with Minimal Media and Lactose as Inducer by Jocksan Ismael Morales-Camacho, Jorge Dominguez-Dominguez, Octavio Paredes-Lopez (61-70).
In this research it was attempted to overexpress the acidic subunit, from the 11S amaranth seed globulin termedamarantin, modified with antihypertensive peptides in Escherichia coli Rosetta (DE3) by manipulating some factors inbatch fermenter such as growth medium composition, inducer (isopropyl ?-D-thiogalactopyranoside [IPTG] or lactose),air flow, cultivation temperature, agitation speed and induction time. The possibility of using several minimal media andlactose as inducer to increase yields of the recombinant protein was investigated. Previous fermentations at flask levelshowed that two minimal culture media (A6 and A7) and 0.5% (w/v) lactose presented high yields of the engineered proteinexpression. Thus, the latter two media were tested at fermenter level, the lactose inducer, and different environmentalconditions. Factors with significant effects were identified by Plackett-Burman design with center points and were adjustedat the level suggested and the yields of the recombinant protein were increased from 303.2 to 1,531 mg L-1 in A6and from 363.4 to 1,681 mg L-1 in A7. Unlike some patents where the highest productivity was achieved at 24 h or afterwards,in this research the best productivity of the recombinant acidic subunit was attained at 4 and 6 h of induction usingboth media, respectively.

A Dual Mode Pulsed Electro-Magnetic Cell Stimulator Produces Acceleration of Myogenic Differentiation by Walter D. Leon-Salas, Hatem Rizk, Chenglin Mo, Noah Weisleder, Leticia Brotto, Eduardo Abreu, Marco Brotto (71-81).
This paper presents the design and test of a dual-mode electric and magnetic biological stimulator (EM-Stim).The stimulator generates pulsing electric and magnetic fields at programmable rates and intensities. While electric andmagnetic stimulators have been reported before, this is the first device that combines both modalities. The ability of thedual stimulation to target bone and muscle tissue simultaneously has the potential to improve the therapeutic treatment ofosteoporosis and sarcopenia. The device is fully programmable, portable and easy to use, and can run from a battery or apower supply. The device can generate magnetic fields of up to 1.6 mT and output voltages of +/- 40 V. The EM-Stim acceleratedmyogenic differentiation of myoblasts into myotubes as evidenced by morphometric, gene expression, and proteincontent analyses. Currently, there are many patents concerned with the application of single electrical or magneticstimulation, but none that combine both simultaneously. However, we applied for and obtained a provisional patent fornew device to fully explore its therapeutic potential in pre-clinical models.