Citoxlab Group – March 2019 Newsletter

Latest news from the Citoxlab Group

High-throughput gene expression profiling and genotyping now available at Citoxlab using the Affymetrix GeneTitan™ Multi-Channel instrument

GeneTitan

Citoxlab further invests to broaden our comprehensive genomics solutions for gene expression profiling and genotyping (Affymetrix cartridges, Next Generation Sequencing and PCR/RT-PCR), with the acquisition of a ThermoFisher / Affymetrix GeneTitan™ Multi-Channel (MC) instrument.

This technology allows streamlining of array processing for discovery, exploration and screening for basic or applied research. The GeneTitan™ MC instrument seamlessly integrates hybridization, washing and imaging in a single instrument to provide automated array processing, allowing highly consistent reproducible data within short turnaround times and all under controlled costs.

In addition to our high quality nucleic acids extraction services from cells, numerous animal and human tissues (including FFPE samples), as well as blood and biological fluids (> 120 000 samples already processed), two new protocols have been recently validated:

  • Gene expression profiling of human, mouse and rat, using the most popular and widely cited microarrays for whole-transcriptome or classical 3’ based designs. This solution offers consistently reproducible data and high accuracy in expression analysis studies.
  • Genotyping, including genome-wide association studies (GWAS), replication studies, candidate-gene association and targeted genotyping for human disease research genomics, using the Axiom™ Genotyping Services.

Like all of our genomics solutions, this new service is available for the analysis of research, preclinical and clinical samples.


Citoxlab and AccelLAB expand their portfolio of preclinical orthopedic surgical models with performance assays adapted to evaluate the latest anti-microbial technologies

orthopedic AccelLAB

Building on a strong imaging and histopathology platform, Citoxlab and AccelLAB are now expanding our orthopedic surgery models to include compromised critical-size defect models and to offer anti-infective efficacy evaluation to the medical device and pharmaceutical industries.

This is in response to orthopedic and trauma device-related infection (ODRI), which remains one of the major complications in modern orthopedics for elective surgeries, with an estimated 3% incidence, along with an increase in absolute case numbers due to the yearly increase in total joint replacements.

Staphylococcus aureus infections account for most chronic osteomyelitis cases, a major challenge in orthopedics. Recurrence of infection in presence of an implant may relate to biofilm-dwelling bacteria persisting deep within cortical bone, with important consequences for patients and also significant financial costs each year. Delivery of antibiotics to the infection site has become a regular adjunct in the treatment of ODRIs. Therapeutic applications in this area are progressing, with a remarkable rate of innovation supported by the rapid evolution of controlled-release formulations and clinical meta-analyses, confirming outcome improvements with strategies employing local delivery of pharmacologically active agents.

With years of expertise in the conduct of orthopedic performance models, our multidisciplinary team combines pharmaco-toxicology and biomaterials experts focusing on the most prevalent device-related infection strain and developing osteomyelitis models for the evaluation of novel anti-microbial coatings and synthetic bone grafting technologies in support of the treatment of implant-associated infections. Our team is now well-positioned to perform these assays in a controlled, Biosafety Level 2 (BSL-2), high technology environment.


High Resolution Mass Spectrometry Sciex TripleTOF® 6600 now available at Atlanbio; research program investigating TOF technology for the bioanalysis of small and large molecules by HRMS

Atlanbio

Atlanbio is pleased to announce the installation of a High Resolution Mass Spectrometer (HRMS) TripleTOF® 6600 (Sciex). This is the first step in a major research program in collaboration with Sciex. This research program will complement our already comprehensive range of bioanalysis services for the toxicokinetic (TK) / pharmacokinetic (PK) evaluation of both small and large molecules.

The development of large molecules is growing within pharmaceutical and biotechnology pipelines, such as peptides, proteins, antibodies, oligonucleotides and many endogenous biomarkers, and their analysis often requires the use of surrogate peptides produced via enzymatic digestion. This last step greatly elevates the complexity of the sample in biological matrices and can produce unwanted interference. HRMS can overcome this lack of specificity encountered when using classical triple-quadrupole technology. Also, HRMS offers a powerful tool for method development, notably for the selection of the peptide with its capacity to provide protein and peptide sequence information as well as modifications (deamidation, oxidation, post-translational modifications). Moreover, its high resolution and extended mass range are well suited for analysis of intact molecules such as monoclonal antibodies and antibody-drug-conjugates, notably for the measurement of the drug-antibody ratio (DAR).

Small molecule bioanalysis routinely performed using a triple-quadrupole are also greatly challenged by endogenous interferences, especially from metabolic compounds. The use of HRMS can increase selectivity and sensitivity, allowing for the possibility of simplifying the sample preparation steps, leading to faster method development and resulting in reduced costs.

Finally, HRMS is a major asset for deeply characterizing reference standards and monitoring compound stability.

 

AccelLAB celebrates 15 years serving the medical device industry by opening a brand-new histology laboratory

AccelLAB new lab

AccelLAB, a Citoxlab company and preclinical contract research organization offering implant safety and performance evaluation to the medical device industry, announces that it is opening a brand-new histology service center with a 3800 sq. ft. fully equipped laboratory.

With this, the preclinical research center has significantly expanded and upgraded its medical device evaluation service with state-of-the-art laboratories including on-site non-decalcified histology embedding/processing centers equipped with two Exakt microgrinding / micropolishing instruments, as well as two Micro Computed Tomography (Micro-CT) instruments (Nikon XT H 225 and a brand-new Bruker Skyscan 1272). All this offered under full GLP compliance with highly qualified on-site technical and pathology staff.

“Scaling up our histopathology service offering by doubling our original histology lab space marks another important milestone reached in our 15-year history,” says Dr. Guy Leclerc, President and CEO of AccelLab. “This expansion will resume in 2019 with major infrastructural changes including additional vivarium suites and operating rooms, which will be implemented later this year”. This reflects AccelLAB’s commitment to remain at the forefront of preclinical and translational research while accelerating device and biologics development programs for faster access to market with turn-key services that have long been recognized for, maintaining the highest industry standards of quality, health, and research practices.


Surge of interest in juvenile minipig studies and milk sampling is driven by recent guidance

Minipig Expertise

With increasing reference to Minipig studies, new regulatory and technical guidance in toxicology is opening up opportunities for Citoxlab, a leader in the use of minipigs in safety testing. In drug development, recent regulations include the much-awaited Step 2 draft of the ICH S11 Guideline on Nonclinical Safety Testing in Support of Development of Paediatric Medicines. Within the field of nutrition, recent guidance from EFSA (European Food Safety Authority) deals with risk assessment of substances present in food intended for infants below the age of 16 weeks. Both guidelines make clear reference to the use of juvenile minipigs in safety testing. In the ICH S11 document, this reference is specifically to Göttingen Minipigs.

The use of juvenile animals in testing is more than just using smaller and younger animals to replace the adult animals used in standard testing. Studies must be designed in ways that are appropriate to the stage of development of the animal, and employing techniques that will ensure the validity of the studies. A factor to consider in juvenile studies is the potential exposure of the juvenile animals to the test item through the mother’s milk. Drug levels must be measured not only in the treated animals (e.g. the mother sow) but also in the milk of the lactating sow and lastly in the blood of the suckling piglets. Skill is needed in the collection of milk samples from lactating minipigs. This technique requires training and also patience! The lactating sows must be separated from the juveniles for a period – normally about 1 hour – before milk collection, in order to ensure that there is sufficient milk to be sampled. We find it beneficial that the technicians performing the sampling procedure are familiar with and socialized with the sows to be sampled. Regardless of the proficiency of a technician, in the absence of previous social contact with the sow, she will not allow herself to be milked. We have invested our efforts in developing the necessary skills to take these samples and generate the needed exposure profiles.

In summary, our experience with Göttingen minipigs, juvenile minipigs and piglets, along with their mothers is important both for our clients, as ever greater use is made of this animal model.

 

Andy Makin appointed as Göttingen Minipig Ambassador

Andy Ambassador

We are proud to announce the nomination of Andrew Makin as Göttingen Minipigs Ambassador, in recognition of his long experience of working with these animals. This year marks the 50 years Jubilee of the founding of Ellegaard Göttingen Minipigs (EGM) and in celebration they will appoint a number of Göttingen Minipigs Ambassadors.  Andy is the first Ambassador to be appointed and he received this honorific title after speaking at a Jubilee Scientific Symposium hosted by EGM in Copenhagen on 28 February. He also received a Certificate of Recognition for his high-level international dissemination of knowledge, promotion of Göttingen Minipigs use in biomedical research and for his contribution to the characterization, validation and development of Göttingen Minipigs disease models. Congratulations Andy!

 

Validation of the Collembolan (Folsomia candida) reproduction test in soil – a new addition to the ecotoxicology panel of tests at Citoxlab

Ecotox expertise

Citoxlab has successfully validated the Collembolan reproduction test in soil (OECD No. 232, 2016, and ISO 11267, 2014) to further expand our ecotoxicology testing portfolio at our Hungarian site. Soil-dwelling Collembola is a relevant species for ecotoxicology testing.

The species has a thin exoskeleton which is highly permeable to air and water; therefore they represent an arthropod species with a different route and rate of exposure as compared to earthworms and enchytraeids.

An appropriate culture method was established to provide age synchronized animals (Folsomia candida springtails) for the test. The test method focuses on the determination of possible effects on the reproduction of springtails by dermal and alimentary uptake in a defined artificial soil substrate. Boric acid, a recommended reference substance, was used as the test item for the method validation study. In the 28-day experiment, the mortality of adults as well as impairment of their reproduction capacity was examined following treatment with several concentrations of boric acid. The observed mortality and reproduction rate of control adults, and the resultant No-Observed-Effect Concentration (NOEC) of boric acid on adult mortality and reproductive capacity were in line with the criteria of the relevant guidelines.

SOLVO remains at the forefront of transporter science with 10 additional publications in 2018, for a total of 85 articles

Transporter expertise

With 2019 well underway, a look back to 2018 reveals that the year was one of the most productive in SOLVO’s history, with a total of 10 scientific articles published. This brings the total number of articles authored by SOLVO scientists to 85, a figure that far exceeds our competitors in the field of drug transporters and transporter-mediated drug-drug interactions, and highlights the company’s dedication to remaining at the forefront of transporter science.

We are truly honored by the number of our colleagues in the transporter field who have chosen to partner with SOLVO for these collaborative programs and look forward to future efforts to help accelerate developments in transporter-mediated ADME-Tox research!

SOLVO Biotechnology publications in 2018:

  • Successful Prediction of In Vivo Hepatobiliary Clearances and Hepatic Concentrations of Rosuvastatin Using Sandwich-Cultured Rat Hepatocytes, Transporter-Expressing Cell Lines, and Quantitative Proteomics. Ishida K, Ullah M, Tóth B, Juhasz V, Unadkat JD.
  • Baicalin is a substrate of OATP2B1 and OATP1B3. Kalapos-Kovács B, Juhász V, Temesszentandrási-Ambrus C, Magda B, Szabó PT, Antal I, Klebovich I, Krajcsi P.
  • Kinetic characterization of bile salt transport by human NTCP (SLC10A1). Jani M, Beéry E, Heslop T, Tóth B, Jagota B, Kis E, Kevin Park B, Krajcsi P, Weaver RJ.
  • Third-generation Sequencing Reveals Extensive Polycistronism and Transcriptional Overlapping in a Baculovirus. Moldován N, Tombácz D, Szűcs A, Csabai Z, Balázs Z, Kis E, Molnár J, Boldogkői Z
  • Transport Kinetics, Selective Inhibition, and Successful Prediction of In Vivo Inhibition of Rat Hepatic Organic Anion Transporting Polypeptides. Ishida K, Ullah M, Tóth B, Juhasz V, Unadkat JD.
  • Human OATP1B1 (SLCO1B1) transports sulfated bile acids and bile salts with particular efficiency. Tóth B, Jani M, Beéry E, Heslop T, Bayliss M, Kitteringham NR, Park BK, Weaver RJ, Krajcsi P.
  • Inhibitor selectivity of CNTs and ENTs. Vaskó B, Juhász V, Tóth B, Kurunczi A, Fekete Z, Zolnerciks JK, Kis E, Magnan R, Bidon-Chanal Badia A, Pastor-Anglada M, Hazai E, Bikadi Z, Fülöp F, Krajcsi P.
  • Correlation Analysis of Potential BCRP Probes in Different Monolayer Systems. Sáfár Z, Jani M, Makai I, Fekete Z, Bui A, Molnár É, Pádár P, Pratt JR, Kis E, Beéry E, Krajcsi P.
  • The oncomir face of microRNA-206: A permanent miR-206 transfection study. Mihály D, Papp G, Mervai Z, Reszegi A, Tátrai P, Szalóki G, Sápi J, Sápi Z
  • Role of efflux transporters in the absorption, distribution and elimination in rodents of a novel PDE4 inhibitor, CHF6001. Cenacchi V, Salvadori M, Riccardi B, Brogin G, Ghiglieri A, Messina M, Imre G, Puccini P