Long-term exposure to polybrominated diphenyl ethers (PBDEs) or polychlorinated biphenyls (PCBs) through diet affects embryonic development of common sole (Solea solea)

Persistent organic pollutants (POPs), such as PCBs (polychlorinated biphenyls) and PBDEs (polybrominated diphenyl ethers), are lipophilic and tend to bioaccumulate in cellular membranes and fat. The accumulation of lipids in oocytes during vitellogenesis, and subsequently in the eggs, favors the accumulation of POPs, and may affect the development of embryos. The effects of dietary exposure to POPs in Common sole Solea solea were evaluated on early life stages (ELS): the survival of sole embryos and features of embryonic development were assessed until hatching endpoint and compared to that of the control. No embryonic development was observed in PBDE eggs while hatching occurred in both control and PCB groups. In addition, several developmental abnormalities were observed in embryos from the PCB groups.Les polluants organiques persistants (POP), tels que les PCB (biphényles polychlorés) et PBDE (polybromodiphényléthers), sont lipophiles et ont tendance à se bioaccumuler dans les membranes cellulaires et les graisses. L'accumulation de lipides dans les ovocytes au cours de la vitellogenèse et par la suite dans les oeufs, favorise l'accumulation des polluants organiques persistants et peut affecter le développement de l'embryon. Les effets de l'exposition alimentaire aux POP chez la sole commune Solea solea ont été évalués aux premiers stades de la vie: la survie des embryons et les caractéristiques du développement embryonnaire ont été évalués jusqu'à l'éclosion en comparaison au témoin. Aucun développement embryonnaire n’a été observé dans les oeufs du groupe PBDE, tandis que l'éclosion a eu lieu dans les groupes témoin et PCB. En outre, de plus nombreux cas d’anomalie ont été observés chez les embryons appartenant aux groupes de PCB

Enhanced CHO Clone Screening: Application of Targeted Locus Amplification and Next‐Generation Sequencing Technologies for Cell Line Development

Early analytical clone screening is important during Chinese hamster ovary (CHO) cell line development of biotherapeutic proteins to select a clonally derived cell line with most favorable stability and product quality. Sensitive sequence confirmation methods using mass spectrometry have limitations in throughput and turnaround time. Next-generation sequencing (NGS) technologies emerged as alternatives for CHO clone analytics. We report an efficient NGS workflow applying the targeted locus amplification (TLA) strategy for genomic screening of antibody expressing CHO clones. In contrast to previously reported RNA sequencing approaches, TLA allows for targeted sequencing of genomic integrated transgenic DNA without prior locus information, robust detection of single-nucleotide variants (SNVs) and transgenic rearrangements. During clone selection, TLA/NGS revealed CHO clones with high-level SNVs within the antibody gene and we report in another case the utility of TLA/NGS to identify rearrangements at transgenic DNA level. We also determined detection limits for SNVs calling and the potential to identify clone contaminations by TLA/NGS. TLA/NGS also allows to identify genetically identical clones. In summary, we demonstrate that TLA/NGS is a robust screening method useful for routine clone analytics during cell line development with the potential to process up to 24 CHO clones in less than 7 workdays

Shortened abstract for Enhanced CHO clone screening using Next-Generation Sequencing and Targeted Locus Amplification publication

Early analytical clone screening is important during Chinese hamster ovary (CHO) cell line development of biotherapeutic proteins to select a clonally derived cell line with most favorable stability and product quality. Sensitive sequence confirmation methods using mass spectrometry have limitations in throughput and turnaround time. Next-generation sequencing (NGS) technologies emerged as alternatives for CHO clone analytics. We report an efficient NGS workflow applying the targeted locus amplification (TLA) strategy for genomic screening of antibody expressing CHO clones. In contrast to previously reported RNA sequencing approaches, TLA allows for targeted sequencing of genomic integrated transgenic DNA without prior locus information, robust detection of single-nucleotide variants (SNVs) and transgenic rearrangements. During clone selection, TLA/NGS revealed CHO clones with high-level SNVs within the antibody gene and we report in another case the utility of TLA/NGS to identify rearrangements at transgenic DNA level. We also determined detection limits for SNVs calling and the potential to identify clone contaminations by TLA/NGS. TLA/NGS also allows to identify genetically identical clones. In summary, we demonstrate that TLA/NGS is a robust screening method useful for routine clone analytics during cell line development with the potential to process up to 24 CHO clones in less than 7 workdays

What is in the fish? Collaborative trial in suspect and non-target screening of organic micropollutants using LC- and GC-HRMS

A collaborative trial involving 16 participants from nine European countries was conducted within the NORMAN network in efforts to harmonise suspect and non-target screening of environmental contaminants in whole fish samples of bream (Abramis brama). Participants were provided with freeze-dried, homogenised fish samples from a contaminated and a reference site, extracts (spiked and non-spiked) and reference sample preparation protocols for liquid chromatography (LC) and gas chromatography (GC) coupled to high resolution mass spectrometry (HRMS). Participants extracted fish samples using their in-house sample preparation method and/or the protocol provided. Participants correctly identified 9-69% of spiked compounds using LC-HRMS and 20-60% of spiked compounds using GC-HRMS. From the contaminated site, suspect screening with participants’ own suspect lists led to putative identification of on average ∼145 and ∼20 unique features per participant using LC-HRMS and GC-HRMS, respectively, while non-target screening identified on average ∼42 and ∼56 unique features per participant using LC-HRMS and GC-HRMS, respectively. Within the same sub-group of sample preparation method, only a few features were identified by at least two participants in suspect screening (16 features using LC-HRMS, 0 features using GC-HRMS) and non-target screening (0 features using LC-HRMS, 2 features using GC-HRMS). The compounds identified had log octanol/water partition coefficient (KOW) values ranging from -9.9 to 16 and mass-to-charge ratio (m/z) of 68 to 761 (LC-HRMS and GC-HRMS). A significant linear trend was found between log KOW and m/z for the GC-HRMS data. Overall, these findings indicate that differences in screening results are mainly due to the data analysis workflows used by different participants. Further work is needed to harmonise the results obtained when applying suspect and non-target screening approaches to environmental biota samples