Cyclic peptides (CPs) constitute a promising biotherapeutics class. Thermal stress studies were conducted to characterize CP stability using multiple biophysical assays, revealing mixed-mode aggregates. New methods were required to decipher the structure of the covalent aggregates. Here, ECD top-down analysis is discussed in the context of mapping CP linkages. The totality of this work gives insights into the in vitro aggregation mechanism, C-C bond lability, and extends disulfide mapping approaches.

Aggregation-induced loss-of-function or oligomerization-induced hyperactivity can be observed with many proteins and removal of aggregated or oligomerized protein is critical for consistent lot-to-lot activity.  SEC, DLS, and MFI analytical testing are all tools used to detect levels of aggregated or oligomerized protein in our purified proteins.  The effects of protein oligomerization on PD-L1/PD1 and SARS-CoV-2 Spike/ACE2 binding properties will be presented along with strategies to mitigate unwanted protein multimerization.

Interface mediated protein aggregation poses a substantial development challenge because it can occur during manufacturing, shipping, storage and/or administration.  In this work, we use state of the art metrology and modeling tools to investigate the mechanism of interface mediated protein aggregation for several monoclonal antibodies and a fusion protein.  We also investigate the impact of intact and degraded polysorbate 80 on this protein degradation pathway.  Improved understanding of protein-surface compatibility could lead to establishing novel and prospective mitigation strategies, spanning protein engineering, formulation development and manufacturing process development.

Talk will address automatic identification of the stress sources of protein aggregates using flow imaging microscopy images, container closure system for suitable for biopharmaceuticals and influence of silicone oil droplets in biopharmaceuticals on immune system. 

Viscosity measurements reflect changes in PPI and the resulting microstructure. Cluster arrangements commonly formed by inherently attractive proteins have a predictable effect on shear rate and temperature dependent viscosity measurements.  We illustrate these effects with model protein solutions formulated with individual amino acid excipients to modify PPI and aggregate structure. 

An early-phase product development shipping study was designed to observe the stability of liquid protein formulations under normal shipping conditions. Shipments were monitored for location, temperature, and shock events. Formulations were evaluated post-shipment for the presence of aggregates and sub-visible and visible particles, and compared against non-shipped and lab-dropped control formulations. The findings demonstrated that factors beyond shock events occurring during shipment are necessary for particle formation in shipped formulations.

The purpose of this presentation is to describe a practical strategy to increase chances of successful translation of promising biologic drug candidates into more stable and commercially feasible drug products.

This presentation will discuss the impact of cool white fluorescent light (CWF) and UV leakage on the mAb aggregation. Surface exposed tryptophan is the major contributor for mAb light sensitivity and this factor was considered for the selection of mAbs based on the number of surface exposed Trp residues. In this study, a concentration dependent CWF light sensitivity is evaluated for different mAbs. The study shows an increase in aggregation with mAb concentration and surface exposed trp residues.

Biologics drug development has experienced rapid growth in recent years. To meet the need, biologics formulation development has quickly acquired a set of automation tools and analytical techniques to provide robust drug products for patients. The push for more effective therapeutics and better patient care options has demanded higher concentrations. This has motivated adaptation of our tools to meet the increases in process complexity for the benefit of patients globally.

Protein biophysical properties are hypothesized to be predictive of protein aggregation under accelerated and long-term storage conditions. Relationship between biophysical properties and protein aggregation is yet to be generalized across a broad selection of mAbs and formulation buffers. We revised historical hypotheses regarding protein physical stability and biophysical properties by combining statistical analysis of the solution behavior of several mAbs mimicking developability and formulation screening workflows.

Protein aggregation and amyloid fibrillation are responsible for several serious pathological conditions like type II diabetes, Alzheimer's and Parkinson's diseases, etc and protein drugs ineffectiveness. Therefore, a molecule that can inhibit the amyloid fibrillation and potentially clear amyloid fibrils is of great therapeutic value. We investigated the anti-amyloidogenic potential of various natural compounds like vitamins, taurine using various biophysical techniques. Aggregation kinetics data, as monitored by ThT fluorescence, inferred that these compounds retards amyloid fibrillation