Assessing Physicochemical Stability of Monoclonal Antibodies in a Simulated Subcutaneous Environment.

Monoclonal antibodies (mAbs) are being increasingly administered by the subcutaneous (SC) route compared to the traditional intravenous route. Despite the growing popularity of the subcutaneous route, our current knowledge regarding the intricate mechanistic changes happening in the formulation after injection in the subcutaneous space, as well as the in vivo stability of administered mAbs, remains quite limited. Changes in the protein environment as it transitions from a stabilized, formulated drug product in an appropriate container closure to the SC tissue environment can drastically impact the structural stability and integrity of the injected protein.

Laser Measurement and Numerical Simulation of Elastomer Stopper Motion during High-Altitude Shipping of Pharmaceutical Syringes.

During high-altitude shipping of pre-filled syringes, pressure differentials can cause the elastomer stopper to move unintentionally. This motion represents a risk to container closure integrity and drug product sterility. To understand and quantitate this risk, we combined high-accuracy laser measurements and numerical simulations of stopper motion.

The journey of a lifetime - development of Pfizer's COVID-19 vaccine.

It would be apt to say that one of the greatest accomplishments in modern medicine has been the development of vaccines against COVID-19, which had paralyzed the entire world for more than a year. Pfizer and BioNTech codeveloped the first COVID-19 vaccine that was granted emergency-use authorization or conditional approval in several regions globally. This article is an attempt to go 'behind-the-scenes' of this development process and highlight key factors that allowed us to move with this unprecedented speed, while adhering to normal vaccine-development requirements to generate the information the regulatory authorities needed to assess the safety and effectiveness of a vaccine to prevent an infectious disease, including quality and manufacturing standards.

The Race to Develop the Pfizer-BioNTech COVID-19 Vaccine: From the Pharmaceutical Scientists' Perspective.

At the outset of the coronavirus disease 2019 (COVID-19) pandemic, it was clear that a vaccine would be crucial for global health efforts. The Pfizer and BioNTech teams came together in a race against the virus, working to design, test, manufacture, and distribute a safe and efficacious vaccine in record time for people around the world. Here, we provide backstory commentary from the pharmaceutical scientist perspective on the challenges and solutions encountered in the development of the Pfizer-BioNTech mRNA COVID-19 vaccine (BNT162b2; b2; Comirnaty®; tozinameran).

Subcutaneous Delivery of High-Dose/Volume Biologics: Current Status and Prospect for Future Advancements.

Subcutaneous (SC) delivery of biologics has traditionally been limited to fluid volumes of 1-2 mL, with recent increases to volumes of about 3 mL. This injection volume limitation poses challenges for high-dose biologics, as these formulations may also require increased solution concentration in many cases, resulting in high viscosities which can affect the stability, manufacturability, and delivery/administration of therapeutic drugs. Currently, there are technologies that can help to overcome these challenges and facilitate the delivery of larger amounts of drug through the SC route.

Accelerating the development of novel technologies and tools for the subcutaneous delivery of biotherapeutics.

Subcutaneous (SC) delivery of biotherapeutics is well established as a route of administration across many therapeutic areas and has been shown to be effective and well-tolerated. It can offer several advantages over intravenous administration. This notwithstanding, there remain critical development issues and knowledge gaps in SC drug delivery.

Transcending the skin barrier to deliver peptides and proteins using active technologies.

Peptides and proteins have been investigated as promising therapeutic agents over the past decade. These macromolecules are conventionally administered by the parenteral route because oral delivery is associated with degradation in the gastrointestinal tract. Transdermal delivery presents a promising alternative route of drug delivery, avoiding pain associated with parenteral administration and degradation issues associated with oral delivery.

Iontophoresis of a 13 kDa protein monitored by subcutaneous microdialysis in vivo.

Background: The purpose of this study was to optimize parameters pertaining to microdialysis technique so as to make this method feasible for evaluating transdermal transport of macromolecules.

Results: Microdialysis experiments were performed in vivo using hairless rats with daniplestim as the model protein. Two perfusion fluids - phosphate-buffered saline (PBS) and 3% dextran in PBS - were evaluated with respect to their effect on sample volume retrieval and recovery of the target protein from the microdialysis probe.

Development of biotechnology products in pre-filled syringes: technical considerations and approaches.

A monoclonal antibody (mAb) product development case study is presented to address some of the issues faced during developing a pre-filled syringe (PFS) product for a biotherapeutic. In particular, issues involving incompatibility with silicone oil and a stability-based approach for selection of PFS barrel and tip cap components have been discussed. Silicone spiking studies followed by exposure to agitation stress or accelerated temperature conditions were used to check for incompatibilities of the mAb with silicone oil, a necessary product contact material in PFS.

Protein and solute distribution in drug substance containers during frozen storage and post-thawing: a tool to understand and define freezing-thawing parameters in biotechnology process development.

Active pharmaceutical ingredient for biotechnology-based drugs, commonly known as drug substance (DS), is often stored frozen for longer shelf-life. Freezing DS enhances stability by slowing down reaction rates that lead to protein instability, minimizes the risk of microbial growth, and eliminates the risk of transport-related stress. High density polyethylene bottles are commonly used for storing monoclonal antibody DS due to good mechanical stress/strain resistant properties even at low temperatures.

Microchannels created by sugar and metal microneedles: characterization by microscopy, macromolecular flux and other techniques.

The objective of this study was to investigate the feasibility of using microneedle technology to enhance transcutaneous permeation of human immunoglobulin G (IgG) across hairless rat skin. Microchannels created by maltose and metal (DermaRoller) microneedles were characterized by techniques such as methylene blue staining, histological examination, and calcein imaging. Methylene blue staining and histological sections of treated skin showed that maltose microneedles and DermaRoller breached the skin barrier by creating microchannels in the skin with an average depth of approximately 150 microm, as imaged by confocal microscopy.

Long- and short-range electrostatic interactions affect the rheology of highly concentrated antibody solutions.

Purpose: To explain the differences in protein-protein interactions (PPI) of concentrated versus dilute formulations of a model antibody.

Methods: High frequency rheological measurements from pH 3.0 to 12.

Transdermal delivery of a approximately 13 kDa protein--an in vivo comparison of physical enhancement methods.

The availability of several enhancement techniques has made it possible to study delivery of macromolecules through skin. This study was conducted to evaluate the transdermal delivery of a ~13 kDa protein using iontophoresis, sonophoresis, and microneedles alone or in combination. In vivo delivery experiments were carried out using hairless rats with daniplestim (DP) as the model protein (molecular weight: 12.

Molecular charge mediated transport of a 13 kD protein across microporated skin.

Transport of proteins across the skin is highly limited owing to their hydrophilic nature and large molecular size. This study was conducted to assess the skin transport abilities of a model protein across hairless rat skin during iontophoresis alone and in combination with microneedles as a function of molecular charge. The effect of microneedle pretreatment on electroosmotic flow was also investigated.

In vitro transdermal delivery of therapeutic antibodies using maltose microneedles.

This paper investigates the microneedle-mediated in vitro transdermal delivery of human IgG as a model protein and demonstrates its applicability to deliver a monoclonal antibody. Microchannels created by the treatment of maltose microneedles in full thickness hairless rat skin were visualized using methylene blue staining. Cryostat sections were prepared and stained using hematoxylin and eosin to locate the depth of penetration.

Ultrasonic rheology of a monoclonal antibody (IgG2) solution: implications for physical stability of proteins in high concentration formulations.

The purpose of this work was to investigate if physical stability of a model monoclonal antibody (IgG(2)), as determined by extent of aggregation, was related to rheology of its solutions. Storage stability of the model protein was assessed at 25 degrees C and 37 degrees C for three months in solutions ranging from pH 4.0 to 9.

Transdermal delivery of interferon alpha-2B using microporation and iontophoresis in hairless rats.

Purpose: To demonstrate transdermal delivery of interferon alpha-2b (IFNalpha2b) in hairless rats through aqueous microchannels (micropores) created in the skin and enhanced by iontophoresis.

Materials And Methods: The Altea Therapeutics PassPort System was configured to form an array of micropores (2.0 cm(2); 72 micropores/cm(2)) on the rat abdomen.

Ultrasonic storage modulus as a novel parameter for analyzing protein-protein interactions in high protein concentration solutions: correlation with static and dynamic light scattering measurements.

The purpose of this work was to establish ultrasonic storage modulus (G') as a novel parameter for characterizing protein-protein interactions (PPI) in high concentration protein solutions. Using an indigenously developed ultrasonic shear rheometer, G' for 20-120 mg/ml solutions of a monoclonal antibody (IgG(2)), between pH 3.0 and 9.

Application of high-frequency rheology measurements for analyzing protein-protein interactions in high protein concentration solutions using a model monoclonal antibody (IgG2).

The purpose of this work was to explore the utilization of high-frequency rheology analysis for assessing protein-protein interactions in high protein concentration solutions. Rheology analysis of a model monoclonal immunoglobulin G2 solutions was conducted on indigenously developed ultrasonic shear rheometer at frequency of 10 MHz. Solutions at pH 9.

Protein structural conformation and not second virial coefficient relates to long-term irreversible aggregation of a monoclonal antibody and ovalbumin in solution.

Purpose: The purpose of the study was to investigate the relationship of the second virial coefficient, B22, to the extent of irreversible protein aggregation upon storage.

Methods: A monoclonal antibody and ovalbumin were incubated at 37 degrees C (3 months) under various solution conditions to monitor the extent of aggregation. The B22 values of these proteins were determined under similar solution conditions by a modified method of flow-mode static light scattering.

Electrically enhanced transdermal delivery of a macromolecule.

The purpose of this study was to establish the delivery parameters for the enhanced transdermal delivery of dextran sulfate (MW 5000 Da). Full-thickness pig skin or epidermis separated from human cadaver skin was used. Silver-silver chloride electrodes were used to deliver the current (0.