18 Jun 2012

Be careful with dust!

While he was walking, he suddenly felt an eye irritation. Thinking that it was just a regular dust, he started to rub his eye, in an effort to remove the dust. Then his eyes got really red, few days passed and his eyes were still red and seem a little swollen. The swelling of his eyes got worse, redder ang bigger. Until he decided to go see a doctor for a check up. The doctor immediately wanted an operation. The doctor thought it was a cyst. Actually, it turned about to be a live worm.
-Janelli Guiang

I just grabbed this from a friend in Facebook. Credits to whoever took this photo. So guys, be careful with dust!

 This is actually my wrst nightmare!! I feel physically sick! :|

5 Jun 2012

Quantitation of Bio surfactant

Biosurfactants might be quantified through surface in addition to interfacial tension. It is a generic quantitation therefore will not identify amongst different types of surfactants which may be present. Biosurfactants may be as opposed in terms of the amount they will reduce surface or maybe interfacial tension, along with the critical micelle concentration (cmc), which can be the minimum surfactant concentration earlier mentioned which not any even more loss of surface tension or interfacial tension takes place.

Measurement of Surface Tension

Surface tension procedures your push vital to relocate some sort of ring engrossed inside a surfactant solution upward as a result of the surface of the liquid into air. Surface tension inside a sample is tested using a surface tensiometer, pro model a Model 21 Tensiomat (Fisher, Pittsburgh, PA ), which works by using the actual du Nouy ring method.

A number of channel elements contained throughout answer, at the grams, sugar, usually do not hinder a du Nouy diamond ring process. Some other channel pieces, for example peptone, and also solvents, for example methanol, could cut down area pressure michael a lack of a new surfactant. For that reason, caution have to be delivered to operate the correct area stress regulates Furthermore, many flying products, for example memory foam or even essential oil, make a difference rating with work surface pressure.

In this instance, the separating could be asked to take away the flying products. However, biosurfactants can be quantified by way of solutions aside from area strain. By way of example, rhamnolipids can be dependant on dimension associated with L-rhamnose because of the orcinol approach.

Parts along with interfacial pressure dimensions are generally dependent upon heat range. Consequently, many examples ought to be equilibrated from space temperatures prior to dimension.

A cmc is dependent upon calculating the top pressure throughout a few examples diluted with 0. 1 Michael, pH 7. 0 phosphate barrier. A normal piece includes record (surfactant focus ) or area pressure and it's utilized to approximate the particular cmc. Understand this image.
Fig. 1. Common piece connected with area pressure up against the record content level connected with mono rhamnolipid. The top pressure appeared to be calculated inside 0. 1 Michael, pH 7 0, phosphate barrier

To look for the surfactant attention in the unidentified test, thin down your small sample before area pressure scored will be preceding the actual minimal area pressure (your surfactant attention with answer will be listed below the actual cmc). A awareness is actually next decided by using a normal piece since proven around number preceding as well as developing by suitable dilution element.

In some instances, a name of any biosurfactant is usually not known along with a basic testing of your raw surfactant answer is usually preferred. Your focus connected with biosurfactant could, consequently, not necessarily become calculated. This sort of primitive surfactant options could possibly be in comparison judging by area pressure.

Dimension of Interfacial Pressure

Interfacial pressure options your pressure recommended to proceed some sort of diamond ring absorbed per fluid, in such cases the hydrocarbon, upwards by having a fluid: Fluid user interface in a 2nd fluid, in this instance drinking water. The process involving finding out surfactant focus inside a test by means of rating with interfacial pressure is just like description involving area pressure.

Ideally by simply reading this article submit a person don’t mistake nowadays around determining the strain regarding biosurfactant plus the focus.

3 Jun 2012

Low Variability ADCC Bioassay

Novel NFAT Reporter Bioassay for Fc Effector Function 

Therapeutic antibodies are now the major class of successful protein drugs with many more in development to target a variety of major diseases. An important and desirable biological activity of most antibody drugs, especially for oncology targets, is antibody-dependent cell-mediated cytotoxicity (ADCC).

In ADCC, circulating natural killer (NK) cells perform lytic killing of antigen-bearing target cells through specific antibody cross-linking of the two cell types. Several signaling pathways are activated in the NK effector cells as a result of multiple FcγRIIIa receptors being engaged by the target cell-bound antibodies. These lead to lysis of target cells and induction of specific cytokines (e.G., through NFAT, or nuclear factor activator of T-cells) in the effector cells.

It is important to quantify the biological activity of any therapeutic antibody in a bioassay during drug development and manufacture. The potency of a manufactured lot is quantified relative to a reference preparation in a lot-release potency bioassay, and this bioassay must be both precise and accurate.

Limits of Classic ADCC Assays

Assays that use primary NK cells possess large inherent variability because of the significant influence of genetic variability and blood donor health status on the extent of ADCC mediated by these cells. Preparation of primary NK cells is slow and tedious. Non-Ab dependent or spontaneous lysis of cells increases background and decreases the dynamic antibody-specific signal range. Extra controls are required to subtract spontaneous lysis from both effector and target cells.

Much current antibody drug development is aimed at establishing new drugs with improved Fc effector function to better address a larger percentage of patients. Accurate and precise quantification of the biological activity of candidate drugs is needed to advance the best candidates through drug development effectively. But this is difficult with current classic ADCC bioassay formats, where inherent variability limits their use in both developmental stages of assessment and manufactured lot-release potency bioassays.

The biologics industry needs better assays, bioassays that can both quantify Fc effector function and that possess less inherent variability and greater ease of use and convenience.

Improved ADCC Assay Design and Procedure

Promega’s reporter-based ADCC bioassay is a new cell-based genetic reporter assay that uses an engineered Jurkat cell line as the effector cell population, avoiding the requirement of purified NK cells from blood donors and avoiding the variability in effector function.

We have exploited activation of the NFAT signaling pathway, which is intact in Jurkat cells and is activated in NK cells upon cross linking of FcγRIIIa receptor with target cell-bound specific antibody. A robust and sensitive luciferase bioluminescence signal generated upon pathway activation delivers good signal to background (fold induction).

We also engineered the same Jurkat cell line to co-express the FcγRIIIa receptor (high affinity variant), thus completing the engineering needed to generate an effector cell line able to quantify Fc effector function of therapeutic antibodies in ADCC. Assay protocol is simple, robust, and convenient. The Promega reporter-based ADCC bioassay is shown in Figure 1.

Cells as Reagents

In our ADCC reporter bioassay, effector cells are essentially assay reagents. The Jurkat effector cells are provided in frozen, thaw-and-use format. They are prepared under stringently controlled conditions for use after thawing directly in the bioassay, without further culture. Ample cell banks enable consistent supply and consistent results. With the improved assay design and procedure, and cells as reagents, the new reporter-based bioassay format outperforms classic ADCC in many key parameters: Low variability, improved accuracy and precision, ease of assay procedure, and low background (sensitivity).

Quantification of Antibody Activity

To demonstrate the ability of the reporter bioassay to quantify Fc effector biological activity of therapeutic antibodies, several on-market antibody drugs were evaluated in the assay. When the anti-CD20 drug Rituximab is assayed using the engineered Jurkat effector cells and CD20+ WIL2-S B cells, clear dose-responsiveness is evident, but when any essential component is absent or replaced with a nonrelevant alternative, e.G., Trastuzumab, no signal is obtained, demonstrating assay specificity (Figure 2A).

The bioassay is effective at quantifying not only biological activity of the therapeutic antibody Rituximab using suspension CD20+ B cell lines as target cells (WIL2-S and Raji) but also biological activity of the therapeutic antibody Trastuzumab using adherent Her2+ cell lines as target cells (SK-BR-3 and MCF7) (Figure 2B).

Using several additional, different biological systems of target cells and antibodies, similar results obtained by alpha testers of this bioassay in the U.S., Europe, and Asia support the “validity” of this bioassay in quantifying the efficiency of Fc effector function in ADCC.

Sensitivity in Detecting Antibody Glycosylation

N-glycosylation of Fc region of antibodies is essential for ADCC. To evaluate the reporter bioassay’s sensitivity to different antibodies with different levels of glycosylation and thus ADCC efficiency, Rituximab was fully deglycosylated using PNGase F. Then blends of N-glycosylated and deglycosylated samples were prepared and assayed in the reporter bioassay (Figure 3). Good sensitivity of the bioassay in differentiating antibodies with small differences in glycosylation is demonstrated by the good linear correlation between the % N-glycosylation and relative activity in the reporter ADCC bioassay compared with fully N-glycosylated antibody.

Bioassay Qualification

Bioassay qualification was performed to evaluate critical performance parameters across a suitable antibody potency range in a potency bioassay. The bioassay demonstrates good linearity, accuracy, precision, and reproducibility from qualification assays run at four different test antibody potencies (range 50-150%) relative to 100% reference sample across three different assay days (data not shown).


Promega’s reporter-based ADCC bioassay provides the biologics industry with a low variability, simple-to-use, and convenient format to quantify the Fc effector function of antibodies in ADCC. It can be widely applied to antibody drug candidates being developed for blood cell and solid tumors, is highly capable of differentiating antibodies with small differences in glycosylation that would affect ADCC efficiency, and has demonstrated good precision and accuracy in bioassay qualification studies

2 Jun 2012



Alternate fuels from renewable cellulosic biomass- plant stalks, trunks, comes, and also leaves-are anticipated to significantly minimize U. S. Reliance upon imported oil even though enhancing national energy security as well as decreasing the environmental effects of energy employ. Ethanol as well as other advanced biofuels from cellulosic biomass is usually environmentally friendly options that can enhance home-based manufacturing regarding travelling fuels, revitalize rural economies, and lessen carbon dioxide as well as pollutant emissions. In line with U. S. Secretary of energy Steven Chu, “Developing another creation involving biofuels will be key to our effort to absolve each of our reliance upon international oil along with address the actual weather situation while generating numerous completely new work of which can’t possibly be outsourced.”
In the United States, the energy Liberty in addition to Security Act (EISA) involving 2007 is an important driver for the ecological progress connected with replenishes able biofuels. In EISA, your Renewable Fuel Common mandates that will 36 billion gallons involving biofuels should be developed every year through 2022, those 16 billion gallons are expected to come by cellulosic feed stocks.
Although cellulosic ethanol output has become demonstrated over a initial level, developing a cost-effective, commercial- scale cellulosic biofuel industry will require transformational science to be able to considerably reduces costs of existing production functions. Woodchips, grasses, cornstalks, and also other cellulosic biomass tend to be widely numerous but tougher to digest straight into carbs when compared with corn grain-the principal source of U. S. Ethanol fuel generation today. Neurological research is actually key for you to speeding up your deconstruction regarding cellulosic biomass straight into sugar that may be converted into biofuels.

The Team of energy (DOE) Office associated with Science continues to play an important purpose in electrifying, helping, along with leading this biotechnology innovation during the last 30 years. The DOE Genomic Science program is improving a new era of analysis centered on achieving whole-systems knowledge of biology. The deal is actually combining professionals throughout diverse fields to understand the particular complicated chemistry and biology root methods to DOE missions inside power generation, ecological remediation, and weather modify science. New interdisciplinary exploration areas are generally emerging, as are knowledgebase along with scientific as well as computational resources essential to be able to developing large-scale, genome-based biology.

Bioenergy Investigation Focuses

To target the most sophisticated biotechnology-based methods for the biological challenges of biofuel manufacturing, DOE recognized three Bioenergy Research Centers (BRCs) inside September 2007. Just about every center is usually chasing the basic analysis underlying numerous high-risk, high-return biological answers pertaining to bioenergy programs? Developments resulting from the actual BRCs provide the knowledge was required to create completely new bio based solutions, strategies, and resources that this rising biofuel industry can use. The particular scientific rationale intended for these kinds of centers as well as regarding other fundamental genomic study crucial towards biofuel business was founded at the DOE class in Biomass to be able to Biofuels regarding customers in the exploration community.

The supreme goal to the three DOE Bioenergy Research Centers is always to far better recognize the actual biological components main biofuel production so that these things is usually redesigned, improved, and helpful to produce novel, effective bioenergy strategies that can be replicated on the mass scale. New methods as well as conclusions emanating from the centers’ essential analysis eventually will certainly advantage just about all biological investigations and definitely will generate the ability actual 3 awesome troubles with the frontiers connected with biology:

Progress connected with next-generation bioenergy crops
Development and layout involving enzymes and microbes together with novel biomass-degrading abilities
Development associated with transformational microbe-mediated approaches for biofuel manufacturing

USDA -- DOE Plant Feedstock Genomics with regard to Bioenergy

DOE and the USDA began any competing scholarship method in 2006 that's invested in essential exploration in biomass genomics, giving your technological groundwork for you to facilitate by using lignocellulose resources with regard to bioenergy and biofuels. Given that lignocellulosic crop plants are generally much less intensive to produce and may grow on lesser top quality land, levels of competition together with crops expanded regarding food manufacturing will be avoided. Observe latest and preceding honors.

More information

Bioenergy Research Centers: An Overview of the Science brochure, July 2010 [faster-download PDF or higher-quality PDF]

DOE Biomass to Biofuels Workshop Summary Breaking the Biological Barriers to Cellulosic Ethanol: A Joint Research Agenda, December 2005, Rockville, Maryland (Order Form)