Molecular Biological Analysis

Molecular biological analyses are used in almost all industries and are an important part of modern laboratory analytics.

Especially in the food industry, molecular biological tests are of great importance, especially when it comes to allergens, genetic engineering and adulteration tests.

Accordingly, the following components can be tested:

• Genetically modified plants
  The most common genetically modified plants include maize, soy and rapeseed. In the food industry, processed products from genetically modified plants such as lecithin and fats are often used.Furthermore, molecular biology plays a major role in feed analysis, because we can also detect genetically modified organisms here. Genetically modified soy meal, for example, is often introduced into animal feed. The laboratories of the Tentamus Group can detect this with the help of molecular biological tests. This is particularly important if the use is undesirable or not even labelled
• Allergens
  According to Regulation (EU) No 1169/2011, there are 14 triggers for allergies or intolerances in the EU.These include:
  1. Gluten
  2. Crustaceans
  3. Eggs
  4. Fish
  5. Peanuts
  6. Soya
  7. Milk
  8. Nuts
  9. Celery
  10. Mustard
  11. Sesame seeds
  12. Sulphur dioxide and Sulphites
  13. Lupins
  14. Molluscs

The Tentamus Group laboratories test your food for the presence of allergens using specific sensitive systems such as PCR, ELISA and enzymatics.

• Certain plants, animal species or viruses
  Furthermore, the laboratories of the Tentamus Group can determine whether certain plants, animal species or viruses are present in a product. For example, it can be detected whether rapeseed is found in mustard. This helps to check processes with regard to cleaning procedures or to avoid cross-contamination.Certain types of fruit and vegetables, such as potatoes, can also be identified.Furthermore, it can be checked whether certain animal species are contained in food products. This is important in the case of religious regulations (e.g. kosher, halal), for example to prove the presence or absence of pork. Food fraud also plays a major role here, as there have been cases in the past where, for example, horse meat was found in lasagne.Contamination with viruses such as plant pathogenic CaMV (cauliflower mosaic virus) can also be detected.Molecular biological analyses are of particular interest for feed, since according to the EU, certain animal components are not allowed to be fed.
• Pathogens
  Another important area of molecular biology is the detection of pathogens such as E.coli. Using molecular biology methods such as PCR, extremely sensitive and rapid pathogen detection is possible. Listeria spp. and Listeria monocytogenes, for example, can be determined in various food groups within 6 hours using RT-PCR.More on the rapid detection of Listeria: “TentaSpeed L. mono”: Testing for Listeria spp. & Listeria monocytogenes in food in 6h!Molecular biological detection of pathogens is also of great importance in animal feed.

In the field of bioanalytics for medical devices and medicinal products, the following molecular biological examination focuses are available:

• Nucleic acid analysis (DNA or RNA)
  The laboratories of the Tentamus Group analyze the identity, content and purity of nucleic acids using conventional or quantitative PCR. This may involve the identity and number of genome copies of, for example, viral vectors used in vaccines as well as microbiological production strains, but also the detection of “model” contaminants such as host cell DNA (host cell DNA), mycobacteria, mycoplasmas and the minute virus of mice (MVM).

  In addition, contaminations by DNA- or RNA-degrading enzymes, so-called DNases and RNases, can be examined in products.

• Functional bioassays
  In the process of functional bioassays or cell effector assays, such as biocompatibility analysis for medical devices (ISO 10993), cell culture-based in vitro techniques are used. Here, products can be tested in various cell culture models for their cytotoxicity, skin irritation and sensitivity. When testing vector vaccines, the viral purity (adventitious agents), the replication capacity and the infectivity of the corresponding batch can be determined, among other things.
• Immune and cell analysis
  Immune and cell analysis covers a wide range of analytical methods, which are carried out according to ICH M3, EMEA, ISO and FDA.

  With the help of ELISA (Enzyme-linked Immunosorbent Assay) systems and flow cytometry (or FACS), various cell- and immunospecific parameters can be analysed. Thus, various cytokines, chemokines and different proteins/enzymes (e.g. host cell protein, benzonase) can be quantified in blood, tissue and production samples.

  In order to map human immuno- and inflammatory reactions (e.g. haemolysis, blood coagulation, complement activation) to medical devices and drugs, ex vivo studies are carried out with fresh human blood.

  In pharmacokinetic (PK) studies, for example, the serum stability of molecules such as antibodies or other analytes is determined over a certain period of time. A special ELISA method on the MesoScale platform (electro-chemo-luminescence) is used for this.

Some areas of the food industry are perceived by the public as particularly sensitive. Here, safety must be ensured through strict quality control.

This concerns in particular:

• GMO-free animal feed

  In order for animal products such as milk to be declared GMO-free, the corresponding feedstuffs must also be free of genetic engineering.

• Vegan and vegetarian products

  There are more and more vegan or vegetarian people. Products labeled as such are subject to strict controls. In particular, products must be checked for animal DNA.

• Food Fraud

  Some foods are susceptible to food fraud. A few years ago, there were frequent cases of horsemeat in lasagne. With the help of molecular biological methods, products can be checked for processing of the desired species in order to detect such substitutions. For example, high-priced fish can be distinguished from lower-priced fish.

  Other products where food fraud frequently occurs are:

  1. Coffee
  2. Tea
  3. Cooking oils
  4. Herbs/spices
  5. Honey
  6. Fruits
  7. Vegetables

  With the help of various methods, including “Next Generation Sequencing” (NGS), food fraud can be reliably detected.

• Adulteration tests (Food Fraud)
  In an adulteration test, certain plants or animal species can be identified. In this way, they can be tested for their presence or absence and adulteration can be prevented.
• Allergen screening
  Products are screened for the 14 substances that trigger allergies or intolerances, as these represent a major health risk for allergy sufferers.
• GMO screening
  The laboratories of the Tentamus Group carry out analytical checks of your food and feed for genetic engineering (GMO/GVO).
• Veggie, vegan and ethical testing
  Test material for the presence of ingredients of animal origin, as well as pork, other animal species and alcohol for halal and kosher products.
• Cytoplasmic male sterility (CMS)
  Our laboratories test for cytoplasmic male sterility, i.e. male infertility in plants, which is based on changes in the DNA. This is prohibited in organic products in Germany and has been found frequently in baby food.
• Detection of microorganisms
  In addition to the classical method, microorganisms can also be detected by molecular biology using PCR. This method is very fast and sensitive in the determination of pathogens.
• Examination of viral vectors
  PCR applications are used to identify introduced transgenes. This supports the production of vaccines and the establishment of gene therapy applications.
• Residual host cell DNA/RNA
  Molecular biological analyses ensure that biopharmaceutical products are free of residual host cell DNA or RNA. The absence of contamination is checked during various production and purification steps. Contamination with nucleic acids can be detected using various PCR and hybridisation methods.
• Applications in clinical trials
  By means of molecular biological analyses, e.g. PCR, relevant parameters for clinical studies can be investigated. To assess pharmacodynamics, enzymatic activities and the expression of specific mRNA, among other things, can be investigated.

For many of our customers, we have already developed new test systems, carried out validation and comparative studies, and provided analytical solutions for specific problems.

PCR

With the help of PCR (polymerase chain reaction) specific nucleic acid sequences can be detected. For example, it is possible to determine which animal species is present in a product or which allergen.

Further information on PCR: Variety identification of apples using PCR

ELISA

ELISA (Enzyme-linked Immunosorbent Assay) is a protein-based method for the detection of larger peptides and proteins. The analysis of the test material is carried out with the help of highly specific antibodies that react with antigens.

NGS

Next Generation Sequencing is a special procedure for DNA sequencing. It can also be used to check mixed products and it is not necessary to know what you are looking for beforehand.

The following laboratories from the Tentamus Group offer molecular biological analyses: