Jen Hug

Recent Posts

Should the Cell Type Affect Your Lysis Method?

Posted by Jen Hug on Oct 30, 2015 12:30:00 PM

should-the-cell-type-affect-your-lysis-methodHave you ever purified protein or cellular organelles? How about extracted DNA or RNA? If you answered yes to any of these, you are probably well-acquainted with at least one cell lysis method. But were you using the optimal method for your specific cell type? Below are listed a variety of cell types matched with the cell lysis method that will derive the highest quality lysate.

Bacterial Cells

In addition to the plasma membrane, bacterial cells are surrounded by a rigid cell wall composed of peptidoglycan. Homogenization is the most commonly used method for lysing bacteria, as it is gentle enough to keep intact the intracellular components but forceful enough to break the cell wall. Laboratories that don’t have access to a homogenizer can alternatively use glass beads (more effective with bacteria than other cell types) or freeze thawing. (1)

Complex Cells

Particularly in large amounts, complex tissue such as muscle and liver requires forceful disruption. These cell types are typically treated with both mechanical lysis and detergent. Mechanical lysis can be accomplished with rotating blades that grind down the tissue. Some models contain an adjustable shaft to accommodate various sample sizes. (2)

Mammalian Cells

Without a tough cell wall, mammalian cells need a method that is effective enough to disrupt their plasma membrane but gentle enough to keep the intracellular contents intact. Therefore, mild detergents with low concentrations of protease inhibitors, or freeze thawing in some cases, are typically sufficient.

Plant Cells

The powerful cell wall comprised of cellulose and polysaccharides makes plant cells difficult to penetrate. Manual lysis via mortar and pestle is therefore one of the quickest and easiest ways to access a plant cell’s intracellular contents.

Small Volumes & Cultured Cells

Cell cultures can be grown with most cell types, mammalian and non-mammalian (e.g. plant, bacteria, yeast). Most cultured cells can be easily disrupted via homogenization; use of a high quality homogenizer may require only 1-2 passes to achieve complete disruption of the sample. Homogenization is also appropriate for small sample sizes or samples that start with small particle sizes. (3)

BEEI: High Quality Homogenizers for Cell Disruption

Most of the cell types listed above can be easily lysed with a homogenizer, regardless of the sample volume. However, the lysate can be of higher quality and more even consistency when run through a top-shelf homogenizer. BEEI is trusted by researchers around the world for both their laboratory homogenizers and their associated customer support.

Cell lysis is just one of a variety of applications for BEEI homogenizers; nano/micro emulsions, lipids, suspensions, and dispersions are also easily achievable. Additionally, the homogenizer processes can be controlled to suit your product, which will allow you to customize to your cell type. And finally, the equipment is easy to use, produces higher yield in less time, and achieves results that are reproducible and scalable.

Learn about how to make your cell lysis protocol more effective by visiting BEEI here or download our FREE eBook on cell lysis below:

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A Complete Guide to E. coli Homogenization

Posted by Jen Hug on Oct 26, 2015 12:30:00 PM

a-complete-guide-to-e.-coli-homogenizationThe E. coli pathogen is responsible for approximately 96,000 foodborne illnesses and $405 million in healthcare costs every year. (1) Yet in spite of its negative image, this bacterial species plays a critical role in laboratories around the world. Specifically, its cells can be ruptured to expose the intracellular contents for numerous applications. Although multiple cell lysis techniques exist, liquid homogenization is the most popular for cultured cells like E. coli. Here is a complete guide to E. coli cell rupture via homogenization, including the equipment you will need, methods and processes, and accurate analysis.

Equipment/Materials

Prior to homogenization, E. coli cells may need to be mixed with a diluting buffer. The optimal buffer depends on the product’s chemical makeup and stability. Once mixed, a cell suspension is ready for homogenization; this process requires appropriate equipment, and there are a variety to select from. For example, ultrasonic, mechanical, and high-pressure homogenization are a few of the more common techniques. In particular, high pressure homogenization allows for reduced sample sizes and uniform consistency. Scientists should have a clear understanding of the specific application they require equipment for before selecting a homogenization method. (2)

Methods & Processes

Although specific methods may change based on the homogenizer and application, most samples will require some form of pre-homogenization treatment and post-homogenization cooling. Many homogenizers, such as the BEE International (BEEI) laboratory homogenizer, can be adjusted to optimize results, particularly across cell types and processes. For example, cavitation, shear, and impact are forces that are critical to successful cell rupture. BEEI homogenizers are structured so that the researcher can adjust these forces to be more gentle or harsh to produce the desired effect. In this way, the optimum results for high yield cell rupture are achieved for the widest variety of cells.

Analysis of Disruption & Particle Size

Analysis of the disruption must take place as close to conclusion of homogenization as possible for an accurate reading. The sample can be viewed with a bright-field or phase-contrast microscope. In additional to its observational strengths, phase-contrast allows the user to gather information on particle size and its impact on downstream processes. The homogenate can then be sedimented to determine protein concentration; these results are to be compared with protein concentrations of prior passes to determine if additional passes are needed. Although a qualitative assessment of particle size can be obtained through a phase-contrast microscope, a quantitative process may be required, specifically in the context of product optimization. Several processes exist to accomplish quantitative analysis, cumulative sedimentation analysis (CSA) is more reliable and requires materials typically found in a laboratory.

BEEI: Homogenizers for Effective Cell Disruption

As your laboratory prepares to run and analyze cell disruptions, you will begin looking at homogenizers that best suit the required needs. BEEI is trusted by pharmaceutical researchers and lab managers around the world. We deliver an array of key benefits, such as production of cell lysates, nano/micro emulsions and dispersions and lipids and suspensions. Our homogenizer processes can be controlled to better suit your product; for example, pressure can be adjusted to be gentler or harsher and the results can be scaled to manufacturing. Finally, our equipment is easy to use, produces higher yield in less time, and results are reproducible and scalable.

Learn more about how BEEI can assist you in cell disruption by visiting us here or for more information, download our FREE eBook below:

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The ABCs of Local VS Systemic Steroids

Posted by Jen Hug on Oct 20, 2015 12:30:00 PM

the-abcs-of-local-vs-systemic-steroidsFor many Americans, the term ‘steroid’ brings to mind famous athletes who were disciplined for its use – Alex Rodriguez, Marion Jones, Lance Armstrong, to name a few. Yet the anabolic steroids used by these individuals are distinct from corticosteroids, which are critical to treatment of a plethora of ailments. Following are the primary modes of corticosteroid delivery and key points about their synthesis. 

Local Delivery

Dermatologic conditions such as psoriasis, eczema, and vitiligo are typically treated with topical corticosteroids (TCS), which work to reduce inflammation in affected skin areas. TCS are intended to affect only the area of application and not the entire system, hence their classification as local and not systemic corticosteroids. TCS are widely used, effective, and easy to apply; the biochemical structure was modeled after naturally-occurring corticosteroids produced in the adrenal gland. (1)

Semisolid preparations, such as topical ointments, oils, and lotions are frequently prepared using mechanical mixers. This mixing action facilitates dispersion, which forms a nice single-phase ointment. From there, homogenization helps to achieve smaller particle size and uniform distribution. (2) Key to this process is use of a high quality homogenizer, such as those produced by BEE International Technologies. For example, their laboratory homogenizers can achieve particle sizes below 100 nm in just one pass.

Systemic Delivery

In contrast to TCS, oral and injectable steroids have the ability to affect all areas of the body, thus their classification as systemic steroids. Yet similar to TCS, systemic steroids are also revered for their anti-inflammatory properties, which can help treat conditions like arthritis, asthma, bronchitis, and colitis. (3) In determining which is a better option- injectable or oral steroid delivery- it may depend on the specific medication in question. However, various misconceptions exist about the ineffectiveness of oral medications, when in fact they can be a more sterile route than injectables; particularly in third-world countries or areas that are unsanitary, injectables carry a higher risk of disease transmission.

Regardless of the delivery method, whether oral or injectable, specific methods must be undertaken to synthesize high quality products. Oral medications in the form of tablets can contain sugar, film, enteric, or compression coatings, which inform the overall tablet composition. As with TCS, oral steroid production requires a homogenizer to achieve the vigorous mixing and granular dissolving required of a high quality product. Use of a high quality homogenizer, such as those from BEEI, can improve attractiveness, ease the swallowing process, and allow for appropropriate internal release. Injectable corticosteroids, distinct from oral delivery, require uniform distribution and small particle size, which can again be achieved via use of a high quality homogenizer.

No matter the delivery method, corticosteroids should be produced with high quality equipment to achieve a consistent and effective product. Homogenization is a key component of the synthesis process; thus, it is in every laboratory’s interest to invest in a high quality homogenizer. BEEI’s laboratory homogenizers can make products such as emulsions, dispersions, lipids, and suspensions, many of which can be used during corticosteroid synthesis.

You can learn more about BEEI’s products here or if you're interested in more information about the particle size reduction process in terms of steroids, download our FREE eBook below:

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4 Things to Ask Yourself Before Selecting a High Pressure Homogenizer

Posted by Jen Hug on Oct 14, 2015 11:30:00 AM

4-things-to-ask-yourself-before-selecting-a-homogenizerSafety and efficacy are the central measurable criteria of any pharmaceutical drug. They may be analyzed by the manufacturer during clinical trials or compared to other drug products by a consumer in the store. Production of a product that will stand out above others requires laboratory equipment that supports synthesis of high-quality product ingredients. In this edition of the BEEI blog, we ask the questions all laboratory managers should ask themselves before purchasing a homogenizer intended for pharmaceutical applications. (1)

  1. Why do I need a homogenizer?

The benefits of homogenizers are numerous, and it is a valuable tool with which to have laboratory access. But before moving into selection criteria, it is vital to ask yourself why you need a homogenizer. Are you conducting R&D and need to disrupt yeast cells? Will you be mass-manufacturing a product that has been approved for market? Some companies, such as BEE International, produce separate homogenizers for each phase of drug development, and models like the Nano DeBEE are better suited for R&D, compared to their manufacture-ready industrial homogenizer.

  1. What motor power does my product require?

This feature is key in that a homogenizer with a higher motor power capacity typically confers a decreased noise level and quicker homogenization. Particularly for researchers who handle high volumes of samples, they should seek out a homogenizer with high motor power capacity for optimal processing efficiency; for example, some motors can be programmed to run up to 1800 watts.

  1. Do I have a desired tube capacity and/or mixing speed?

In other words, how many tubes need to be mixed simultaneously and what speed do they need to be mixed at? This factor may be relevant for laboratories that are producing high volumes of a single product, such as in the latter stages of clinical trials or in manufacturing. Additionally, it will be important to prioritize a high mixing speed for applications requiring high speed homogenization.

  1. Does my product require a specific particle size?

Many products, particularly pharmaceuticals such as vaccines, inhalants, targeted drug delivery, and anesthetics, require consistency in texture and size to achieve safety and efficacy. If your laboratory is synthesizing a product that requires particle sizes at or below 100 µm, you will be able to narrow down your field of homogenizers. Many high pressure homogenizers can produce emulsions, dispersions, and suspensions at particle sizes of 100 µm or higher, yet a select few can achieve down to 0.1 µm. The Micro DeBEE from BEE International, for example, can achieve 0.1 µm and would be an appropriate selection for a product with this specification.

If you are able to identify even one of the above criteria as a priority, you will be able conduct your homogenizer search in a more focused manner; this should optimally yield a purchase that provides maximum laboratory output.

Begin your search by comparing the laboratory homogenizers from BEE International Technology. Our products are trusted by pharmaceutical researchers and lab managers around the world for delivering an array of key benefits; these include production of nano/micro emulsions, dispersions, lipids, and suspensions. In addition, we have extensive experience in the challenges that our pharmaceutical customers face as they transition from concept, through to R&D, clinical trials, all-important FDA approval and finally, to manufacturing.

Learn more by visiting http://www.beei.com/industry/pharmaceutical-process-equipment.

 

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Pharmaceutical Clinical Trials: Demystifying the Process

Posted by Jen Hug on Oct 9, 2015 11:30:00 AM

pharmaceutical-clinical-trials-demystifying-the-processThis is Part 2 of a 3-Part series detailing a product’s pathway from research & development (R&D) to clinical trials to manufacturing.

Of the thousands of promising drugs that enter the R&D phase of pharmaceutical drug development, only a small fraction move forward into clinical trials. And 7 out of 8 drugs that do make it to clinical testing will never reach the marketing phase. (1) The current state of pharmaceutical testing is clearly bleak; researchers and corporations should therefore be informed about each part of the process to increase their likelihood of success. Following are descriptions of each clinical phase and information on how a drug can progress into FDA approval.

IND Application

Before researchers can embark on human drug testing via clinical trials, an Investigational New Drug (IND) application must be filed with the FDA. The FDA requires certain information to be contained within the application; these include 1) pre-clinical study results, 2) manufacturing information, 3) clinical protocols, and 4) investigator information. (2)

Phase I

Upon approval of its IND application, a drug enters Phase I of clinical development. The overarching purpose of Phase I is to assess a drug’s safety as it is administered to humans for the first time. During this time, researchers study measures such as dosage limits, mechanism of absorption, and overall body response; a small human sample size (anywhere from 10-20 people) is typically used in case of adverse side effects. (3)

Phase II

Successful completion of Phase I indicates that a safe drug dose has been identified and it is now ready for effectiveness testing. Using either a single dose or varying doses across treatment groups, researchers administer the drug to a larger group of people (anywhere from 20-40 people). Individuals with diseases that aren’t responding to other forms of treatment often volunteer for Phase II clinical trials in hopes that the experimental drug will effectively treat their condition; experimental cancer drugs are a common example of such practices. (4)

Phase III

An experimental drug is greenlighted into Phase III after it has demonstrated safety and effectiveness in Phases I and II. The purpose of Phase III is to confirm these criteria in larger groups of people; principal investigators may enroll hundreds to thousands of people for a Phase III clinical trial. Most drugs that enter this phase are intended for disease treatment, and may be administered alongside an already existing standard treatment to determine if the new drug is more effective. Phase III may last for years, and upon completion the pharmaceutical company can submit a request to the FDA for its approval. (5)

As you press forward with a potentially promising drug therapy, keep in mind that very few drugs actually make it to the clinical trial stage of drug development. To optimize your drug’s chances of success, it is imperative that your R&D phase yields a high quality drug, which can only be produced with high quality equipment.

At BEEI International, we produces homogenizers that can yield a variety of relevant products, such as emulsions, suspensions, dispersions and lipids. Importantly, these products go on to make up the injectables, inhalants, anesthetics, vaccinations, and more that are used in the clinical trials described here. In addition, we have extensive experience assisting our product users as they transition through the drug development process.

Visit us here to learn more about how their homogenizers can optimize your drug’s chances of success.

Check out Part I of the drug development series for more on the R&D process; and stay tuned for Part III on drug product manufacturing, the final installation of our drug development series!

Particle size reduction equipment: is the microemulsion the drug delivery vehicle of the future?

Posted by Jen Hug on Aug 18, 2014 5:26:00 PM

Delivering therapeutic compounds into people or animals can be quite challenging. Digestive fluids destroy many compounds, including the biologics many pharmaceutical companies have been investigating lately. Taking drugs by injection doesn't appeal to most people, and increases the risk of infection from needles. 

 

Transdermal_patch_for_particle_size_reductionTransdermal drug delivery

One of the major functions of the skin is to keep out foreign agents. The skin does this with great efficiency, including keeping out pharmaceuticals. There are many advantages then, if the skin barrier can be overcome by transdermal delivery of pharmaceuticals. The drug can be released slowly and continuously into the system. Since the drug doesn't enter the body via the digestive tract it doesn't go through the liver's detoxification systems before reaching the parts of the body that need it. The only problem is getting the drug through the skin barrier.

What is a microemulsion?

Microemulsions have been under extensive study recently, as possible transdermal drug delivery systems. A microemulsion is a clear or transparent system with stable particles smaller than 150 nm. The emulsions which most of us are familiar with are cloudy or milky with large particles, and are not stable, eventually separating into its various phases. 

Microemulsions are ideal for transdermal drug delivery. Once created, they are stable. They can carry both hydrophilic and lipophilic drugs. Due to their structure, microemulsions can carry very high concentrations of drugs, and have an enhanced ability to pass through the skin. The small size of the particles, action of the surfactants on the skin, and the continuously fluctuating interphases can breach the skin's barrier.

Studies of the ideal microemulsion to carry specific drugs into the body through the skin are ongoing. Reducing the particle size may improve the penetration of most emulsions. Varying the oils, surfactants, and viscosity of the microemulsion can all affect the ability of the microemulsion to deliver drugs through the skin. Microemulsions can even be created that only deliver drugs into the skin rather than through it. The cosmetic industry uses microemulsions to deliver anti-wrinkle products into the skin's layers. 

Skin irritation?

One concern with the use of microemulsions to deliver drugs is their ability to irritate skin. In tests conducted to date, microemulsions, in general, seem to be far less irritating than solutions of sodium lauryl sulfate, a moderate to severe skin irritant. Most microemulsions have been about as irritating to the skin as saline. Test subjects exposed to microemulsions for up to four days exhibited no skin redness or apparent irritation. 

Microemulsions show great promise as drug delivery systems. If your research lab needs particle size-reduction equipment to expedite research into the ideal microemulsion to carry a particular pharmaceutical across or into the skin, don't hesitate to contact us

2 Ways High Pressure Pasteurization is Utilized in the Food Industry

Posted by Jen Hug on Dec 21, 2013 8:58:00 AM

describe the image2 Ways High Pressure Pasteurization is Utilized in the Food Industry

The food industry has struggled with one question for years, “How do you remove harmful bacteria from food without sacrificing taste and quality?” High pressure pasteurization (or processing; HPP) is becoming a popular choice amongst food manufacturers, due to the fact that it does not expose food to detrimental processing.  Heat, which is a popular choice, could potentially change the flavor and nutritional content of a product. This happens when organic matter has been burnt off while neutralizing bacteria and must be supplemented, resulting in obvious changes in composition.  New ways to pasteurize with high pressure are likely being developed, but at this time there are only two popular methods of production.

Pasteurization using large tanks is fairly common and simple. Prepackaged food is placed into a tank filled with water. That water is then pressurized, meaning that high pressure is equally distributed throughout the tank, neutralizing the bacteria. The food is unchanged because the pressure is equally distributed throughout the food. The advantage of the tank method is that it can pasteurize large amounts of solid food at one time. One downside is getting the food in and out of the tanks takes time and involves manual labor.

The other form of high pressure pasteurization involves using a high pressure homogenizer to pasteurize liquids or semi-solid products. The product is fed directly into the device and then pushed through a system of tubes and nozzles using the intensified pressure generated by a hydraulic pumping system. The advantage to this form of high pressure pasteurization is that it also allows the product to be emulsified or even broken down while being pasteurized. This removes a step for those products that benefit from the combination of cell rupture and organic matter size reduction. While heat could be a factor during HPP (since it is released when pressure is heightened), heat exchangers and chillers can be used to keep product temperatures low, eliminating any harmful side effects from the processing.

Overall, high pressure pasteurization is a profitable option for the food industry. It neutralizes bacteria while leaving taste behind. The option that any manufacturer chooses is going to be based on their needs. For liquid and semi-solid products, using a high pressure homogenizer is both cost effective and recommended. Product can be both processed and pasteurized simultaneously, meaning that much of the labor used during manufacturing can be applied elsewhere and workflow can be streamlined. For solid products, and those that would not benefit from homogenization or mixing, the tank option is perfect. Large amounts of food can be pasteurized at once, after it has been processed, meaning that quality and taste is retained and any harmful bacteria neutralized. No matter which method is appropriate for a product, you cannot go wrong with high pressure. 

photo credit: fotoliene via photopin cc

2 Ways High Pressure is Changing the Pharmaceutical Industry

Posted by Jen Hug on Jul 12, 2013 2:02:00 PM

2 Ways High Pressure is Changing the Pharmaceutical Industry



high pressure homogenizers for pharmaceutical industryWhen we see the words “high pressure” we generally picture ourselves or those around us under distress. Jobs with strict deadlines and high expectations can be emotionally, physically and socially problematic for those that have them.  In addition, a powerful buildup of gases is considered dangerous and could potentially lead to an explosion. Despite the negative connotations associated with them, anyone who pays close enough attention to the weather forecast knows that those words indicate sunny days to come. High pressure forces can also lead to positive results in the pharmaceutical industry by helping to alleviate two major obstacles in the development of new medications.

 

One difficulty the pharmaceutical industry faces is bioavailability. Bioavailability refers to the amount of medication your body absorbs. When a drug is taken orally, the absorption can be much lower than other routes of administration (e.g. nasal, intravenous and epidural), which is problematic since many medications are delivered this way. Even when you take a vitamin, you do not receive the full dosage that you swallow. In drugs with poor water solubility, this bioavailability decreases because the particles are too big to dissolve into water (and water passes easily through the body). Using high pressure machinery to reduce particle size can increase bioavailability and dissolution. For the pharmaceutical industry this means producing medications that have more effective dosages.

 

Another problem the pharmaceutical industry faces is poor cell disruption. As you know, cells are the building blocks of life. Sometimes what is contained within a cell can be valuable, but cell walls can act as a strong defense against the breakdown of that cell.  High pressure forces can create the necessary cell disruption, allowing for the usually unattainable contents to be harvested. While cell disruption can be useful to a number of industries, in pharmacology it can mean creating newer and more effective medications.

 

I know you’re wondering, “How does this affect me? Why should I care?” The answer is simple: high pressure modified medication can improve your quality of life. Prescription drugs that can more effectively absorb into your body mean spending less time sick and more time active. Subsequently, this leads to happier people and since happier people tend to be healthy people, the use of such medications could indirectly lead to longer periods of uninterrupted good health.                                                                

 

So, while high pressure may seem daunting in some contexts, for the pharmaceutical industry it means innovation and improvement. Since high pressure machinery is currently in use in research and development laboratories across the globe, it is only a matter of time before the greater effects of this technology is felt.

 

photo credit: nima; hopographer via photopin cc