Studying protein polymerization with mass photometry

The COVID-19 pandemic has brought global awareness to the dangers of emerging pathogens to human health and welfare. Common molecular tools used at scale, such as PCR, are best suited for detecting known pathogens. Preparing humanity against future zoonotic pandemics requires unbiased surveillance tools, such as next generation sequencing (NGS). This approach enables the unbiased detection of viral sequences (known and unknown), identifies possible co-infections, and provides insights into host transcriptional response. This webinar will introduce a CRISPR-based strategy improving sequence complexity and discovery in libraries prepared from complex biological samples. The critical component enabling this approach is Jumpcode Genomics’ CRISPRclean technology which removes abundant mammalian and bacterial ribosomal RNA sequences from NGS libraries. We will review data from >40 nasopharyngeal samples collected from SARS-CoV-2 testing, demonstrating >90% depletion of all human and bacterial ribosomal sequences, 3-to-7-fold increase in viral and bacterial transcriptome coverage and 5-to-10-fold increase in coverage of mid to low expressing host transcripts.

As the number of biotherapeutic candidates in development is increasing, the requirements of analytical tools to understand the molecules better early in the process are rising, simultaneously. To understand product quality attributes (PQAs) fully, identification, localization, and relative quantification are key. Mass spectrometry (MS) plays an important role with peptide mapping being the gold standard to assess post-translational modifications (PTM). However, traditionally used collision-induced dissociation (CID) has a number of limitations in PTM identification and localization. Here, the novel fragmentation technique electron activated dissociation (EAD) was employed for peptide mapping of a multispecific monoclonal antibody (mAb) as well as IgG1 and IgG4 mAb samples.

In a constant battle to find more efficient and effective ways to test oncology preclinical candidates, 3D in vitro culture methods for PDX-derived tumor cells have shown promise of in vivo-like growth characteristics, invasion, and responses to therapeutics. With these methods, we can perform high-throughput in vitro PDX screening that identifies active and selective molecules, and select favorable PDX tumor models for subsequent validation of candidates in vivo.

Scientist must look beyond studying individual proteins and examine the structure and assembly of protein complexes within cells in order to truly understand how proteins function in their native environment.