Luminex xMAP®: Enhanced lab efficiency

Stereotypical depictions of researchers involve crazy hair, oversized goggles, shabby lab coats, and loads of test tubes. While the first three may be exaggerated, the sheer volume of tubes and wells needed in a lab cannot be overstated, especially when the lab is dedicated to anything biological.

One tissue sample can be used for a gamut of tests, all of them attempting to identify something different in it, be they antibodies, DNA, or RNA (biomarkers). Often, many samples are required due to all the tests needed to highlight the variations in those biomarkers. But the size of samples is now decreasing thanks to machines like the Luminex System running xMAP technology.

The Luminex System is a research/clinical diagnostics platform that allows detection of multiple analytes in a single well of a microtiter plate—100 or more reactions using a single drop of fluid.

Multiplex assays are widely used in experiments investigating the characteristics of molecules within a biological sample. This approach can be used to see whether an experimental treatment works, or what changes a DNA mutation causes in the molecules or molecular pathways within cells.

In real terms, this machine allows for analyses to be done to determine whether or not a patient has a particular disease or gene variant in their blood that would prevent a drug from being effective. It also allows them to determine the ideal dosage for those drugs. The machine can also be used to identify and characterise viral infections.

A particular research group at the University of Malta, headed by Prof. Godfrey Grech, has used Luminex xMAP technology to develop novel markers which are allowing them to classify a subset of triple-negative breast cancer
patients.

By identifying these biomarkers, it may be possible in future to detect the disease earlier and give patients better-targeted therapy.

Prof. Godfrey Grech and his team of researchers.

Author: Prof. Godfrey Grech

Malta’s brightest exports: Travelling to the EU’s JRC

A group of Maltese researchers travel to the European Commission’s Joint Research Centre site in Ispra to share their work in the fields of climate change, environment, and medicine. Cassi Camilleri writes.

Continue reading

Blood, Genes, and You

Over the course of nine months, an entire human body is sculpted from a few cells into a baby. The blueprint is the information written into our DNA. But what happens if there is a mistake in these blueprints? Decades worth of research carried out in Malta and abroad have aimed to understand how these errors lead to a disease common in Malta and prevalent worldwide.
Scott Wilcockson talks to Dr Joseph Borg (Faculty of Health Sciences, University of Malta) to find out more.

Continue reading

Who owns you?

One fifth of human genes have already been claimed as US Intellectual Property. But should anyone own our genes? And what happens when gene ownership can drastically prevent the advancement of life-saving cures?

The US Patent Office’s most controversial patents are on BRCA1 and BRCA2, both linked to the high risks of ovarian and breast cancer. They are now owned by Myriad Genetic Laboratories. In 1996, Myriad Genetics developed and began marketing a predictive test for the presence of possible cancer-causing mutations: the ‘BRCAnalysis’ test. The price of the test was US$3,000 but the company promised that it would eventually drop the price to US$300. This never happened because its patent holder had the right to stop any other party from duplicating the patented sequences. This single test accounted for over 80% of Myriad Genetics’ multibillion dollar business.

In 2009, the American Civil Liberties Union (ACLU) decided to challenge the patenting of human genes on legal grounds. The ACLU was the representative of 20 medial organisations, geneticists, women’s health groups, and patients unable to be screened due to the prohibitive patents. The ACLU’s position was that Myriad’s patents violated the patent law on the issue of patent-eligibility.

The case went before the Supreme Court. By 3 June, 2013 it was declared that the Myriad patents were invalid because they did not create or alter any of the genetic information encoded in the BRCA1 and BRCA2 genes. The location and order of the nucleotides existed in nature before Myriad found them. The company simply discovered what was already there and did not create anything new.

There is no worldwide consensus on whether parts of the human genome should be granted intellectual property protection. The Myriad patents should alert us to the injustice of having a pharmaceutical company make money out of cancer predictive tests that could cost 10 times less than what is charged. The same patents stifled diagnostic testing and research that could have led to cures as well as limiting women’s options regarding their medical care in Malta as in all other parts of the world. There are various international and regional agreements that have described the human genome as being part of humanity’s ‘common heritage’, including the 1998 UN Declaration on the Human Genome and Human Rights. The Myriad patents controversy has shown that gene patenting does not work to stimulate more research—one of the prime arguments Big Pharma uses. It is time to explore other avenues that will both promote scientific progress and technological development but at the same time protect the special nature of human genes that make us who we are. No one should own our genes—they should be exploited in the interest of everyone.

Written by: Dr Jean Buttigieg

The Hidden History of the Maltese Genome

By reading someone’s DNA one can tell how likely they are to develop a disease or whether they are related to the person sitting next to them. By reading a nation’s DNA one can understand why a population is more likely to develop a disease or how a population came to exist. Scott Wilcockson talks to Prof. Alex Felice, Dr Joseph Borg, and Clint Mizzi (University of Malta) about their latest project that aims to sequence the Maltese genome and what it might reveal about the origins and health of the Maltese people.

Continue reading

Heartbreakers

Every person possesses the same genes within every cell. Their DNA provides the information to first create an entire functioning body and then keep it running. While all humans share more than 99.9% of their DNA, it is the subtle differences in our DNA that ensure individuality. Many differences are superficial effects, like hair colour, but some can have disastrous health effects. Scott Wilcockson talks to Dr Stephanie Bezzina-Wettinger (Faculty of Health Sciences, University of Malta) about her research on these subtle differences and how they can contribute to heart attacks.

Continue reading

Maltese Olives and their genes

The olive tree (Olea europaea L.) is one of the oldest species of domesticated trees and the second most important oil fruit crop cultivated worldwide. 97% of the global olive cultivation is concentrated in the Mediterranean Basin. The olive thrives in Maltese soils. Economically, olives are not important for local agriculture, but its cultivation is becoming popular since the Maltese agribusiness has a lot of room for growth to make high quality oil and secondary products. 

Bajda-fruit4-RecoveredIn the Mediterranean region there are two subspecies of Olive tree. These are the wild olive (O. europaea L. subsp. Oleaster) and the cultivated olive (O. europaea L. subsp. Sativa). Each subspecies has several cultivars selected for taste, size, disease resistance or other desirable qualities. There are 1,300 cultivars worldwide and Malta is no exception. The Maltija cultivar is probably the most popular Maltese cultivar and can give a high productivity. The Bidnija cultivar, which is believed to be the oldest Maltese olive cultivar (it is thought to date back to Roman times), produces oil of excellent quality rich in polyphenols (these have many health benefits), exhibits high tolerance to environmental stress such as salinity and drought, and demonstrates resistance to pathogens and pests such as the olive fruit fly. The Bajda variety produces a characteristic white drupe. Besides the native cultivars, there are a number of Maltese wild olives. 

Renowned foreign varieties associated with high productivity tend to have a higher productivity than local cultivars. For this reason, local farmers find foreign varieties more convenient, leaving Malta at risk of forever losing its unique olives.

Till now revival efforts focus on artificial propagation and re-plantation. These trees are identified by their appearance. This is an inaccurate method since olive growth is influenced by environmental conditions.Bidni-fruit-+-leaves

To develop a better way to identify local cultivars, Oriana Mazzitelli (supervised by Dr Marion Zammit Mangion) has focused on adopting a genetic approach. She also wanted to examine the genetic diversity of Maltese olive varieties. Mazzitelli compared the genetic patterns of local varieties to those generated by two commercial Italian (Carolea) and Tunisian varieties (Chemlali). The genetic analysis produced unique DNA profiles that can provide a more accurate means of identification than just looking at the plant.

The genetic variability between varieties was high. The Bidnija and Maltija stood out for their genetic uniqueness. The differences between local varieties suggest that, despite being allegedly native, the origins of the two are not directly linked. A number of DNA marker regions detected in the foreign cultivars and in the Maltese wild olive were undetected in the Maltese cultivars, suggesting that not all DNA markers are present and amplifiable in foreign varieties have been conserved in the Maltese cultivars. Mazzitelli’s work is an important first step to show that local varieties can be identified cheaply through DNA analysis. Without genetic identification, maintaining and cultivating local varieties would be near impossible—a case of genes for good olive oil.

 

This research is part of a Master of Science in Biochemistry at the Faculty of Medicine and Surgery, University of Malta. The research was funded by STEPS (Strategic Educational Pathways) scholarship which is part-financed by the EU’s European Social Fund (ESF) under Operational Programme II—Cohesion Policy 2007-2013, ‘Empowering People for More Jobs and a Better Quality of Life’. 

 

Developing new cancer treatments

The lab is my second home, with the rugby pitch a close third. My fascination with lab work and science started when I visited Tays Hospital in Finland. It was during my bachelor degree in Medical Laboratory Science. This three-month placement helped me choose cytogenetics for my final year project. My work involved developing a technique to allow for doctors to better manage sporadic and recurrent miscarriage patients.

My interest in cytogenetics (the study of chromosomes where genes are found) evolved to genetics, when I started working at the biotech company MLS BioDNA Ltd. This laboratory focused on the testing of inherited diseases, paternity and forensics, as well as food and water microbiology. Working in a diagnostic laboratory was very satisfying but I had always wanted to pursue research. So I moved to Sheffield to read for a Masters in Molecular Medicine, with the help of the Malta Government Scholarship Postgraduate Scheme (MGSS). My intention was to just stay for the course and return home, however, my current supervisor offered me a 10-month contract to work in a molecular microbiology lab. This was a very pleasant experience, and encouraged me to pursue a Ph.D. I received a scholarship for a Ph.D. in Immunology at the University of Sheffield, which I am currently working on.

Vaccines can prevent certain infectious diseases. Potentially, they can also treat cancer. Vaccines today are based on small proteins, which by themselves do not elicit a strong immune response. To treat cancer a strong response is needed. Immunological adjuvants that amplify the immune response are used to accomplish this. However, no one really understands how these adjuvants work. For my Ph.D., I am part of a research group that focuses on an immunological adjuvant which increases the immune response by over 1,000 times. Understanding how these adjuvants work will pave the way to more targeted treatments and fewer side effects.
My job is to understand which immune cells are responsible for this effect. The adjuvant has been shown pre-clinically to be effective in B cell lymphoma, a type of cancer of the blood that originates in the lymph glands. Patients are currently treated with the drug Rituximab which depletes certain immune cells called B cells. If our treatment requires other immune cells to work, it can be used in addition to therapies such as Rituximab.

Although a Ph.D. is something which I really wanted to do, it was still a shock to my system. Scientific research can be very frustrating as long hours and hard work do not necessarily translate into results. In spite of this, the long-term goal of this project keeps me going making the sweat and tears worth it.