Using X-Ray Crystallography to Simplify and Accelerate Biologics Drug Development

Every major biopharmaceutical company incorporates a protein crystallography unit that is central to its structure-based drug discovery efforts. Yet these capabilities are rarely leveraged toward the formal higher order structural characterization that is so challenging but integral to large-scale biologics manufacturing. Although the biotech industry laments the shortcomings of its favored biophysical techniques, x-ray crystallography is not even considered for drug development. Why not? We suggest that this is due, at least in part, to outdated thinking (for a recent industry-wide survey, see Gabrielson JP, Weiss IV WF. Technical decision-making with higher order structure data: starting a new dialogue. J Pharm Sci. 2015;104(4):1240-1245). We examine some myths surrounding protein crystallography and highlight the inherent properties of protein crystals (molecular identity, biochemical purity, conformational uniformity, and macromolecular crowding) as having practicable commonalities with today\u27s patient-focused liquid drug products. In the new millennium, protein crystallography has become essentially a routine analytical test. Its application may aid the identification of better candidate molecules that are more amenable to high-concentration processing, formulation, and analysis thereby helping to make biologics drug development quicker, simpler, and cheaper

Encapsulation state of messenger RNA inside lipid nanoparticles.

Understanding the structure of messenger RNA (mRNA) lipid nanoparticles, and specifically the microenvironment of the mRNA molecules within these entities, is fundamental to advancing their biomedical potential. Here, we show that a permeating cationic dye, thionine, can serve as a cryogenic electron microscopy contrasting agent by binding selectively to encapsulated mRNA without disturbing lipid nanoparticle morphology. Cryo-electron microscopy images identify the mRNA location, revealing that mRNA may exist within solvent-filled cavities or may be substantially lipid associated

Recommended C Style and Coding Standards

This document is an updated version of the Indian Hill C Style and Coding Standards paper, with modifications by the last three authors. It describes a recommended coding standard for C programs. The scope is coding style, not functional organization. July 4, 1990 Recommended C Style and Coding Standards L.W. Cannon R.A. Elliott L.W. Kirchhoff J.H. Miller J.M. Milner R.W. Mitze E.P. Schan N.O. Whittington Bell Labs Henry Spencer Zoology Computer Systems University of Toronto David Keppel EECS, UC Berkeley CS&E, University of Washington Mark Brader SoftQuad Incorporated Toronto 1. Introduction This document is a modified version of a document from a committee formed at AT&T's Indian Hill labs to establish a common set of coding standards and recommendations for the Indian Hill community. The scope of this work is C coding style. Good style should encourage consistent layout, improve portability, and reduce errors. This work does not cover functional organization..

Recommended C Style and Coding Standards

This document is an updated version of the Indian Hill C Style and Coding Standards paper, with modifications by the last three authors. It describes a recommended coding standard for C programs. The scope is coding style, not functional organization

Examination of Thermal Unfolding and Aggregation Profiles of a Series of Developable Therapeutic Monoclonal Antibodies

Screening for pharmaceutically viable stability from measurements of thermally induced protein unfolding and short-term accelerated stress underpins much molecule design, selection, and formulation in the pharmaceutical biotechnology industry. However, the interrelationships among intrinsic protein conformational stability, thermal denaturation, and pharmaceutical stability are complex. There are few publications in which predictions from thermal unfolding-based screening methods are examined together with pharmaceutically relevant long-term storage stability performance. We have studied eight developable therapeutic IgG molecules under solution conditions optimized for large-scale commercial production and delivery. Thermal unfolding profiles were characterized by differential scanning calorimetry (DSC) and intrinsic fluorescence recorded simultaneously with static light scattering (SLS). These molecules exhibit a variety of thermal unfolding profiles under common reference buffer conditions and under individually optimized formulation conditions. Aggregation profiles by SE-HPLC and bioactivity upon long-term storage at 5, 25, and 40 °C establish that IgG molecules possessing a relatively wide range of conformational stabilities and thermal unfolding profiles can be formulated to achieve pharmaceutically stable drug products. Our data suggest that a formulation design strategy that increases the thermal unfolding temperature of the Fab transition may be a better general approach to improving pharmaceutical storage stability than one focused on increasing <i>T</i>_onset or <i>T</i>_m of the first unfolding transition

Recommended C Style and Coding Standards

This document is an updated version of the Indian Hill C Style and Coding Standards paper, with modifications by the last three authors. It describes a recommended coding standard for C programs. The scope is coding style, not functional organization. February 19, Recommended C Style and Coding Standards L.W. Cannon R.A. Elliott L.W. Kirchhoff J.H. Miller J.M. Milner R.W. Mitze E.P. Schan N.O. Whittington Bell Labs Henry Spencer Zoology Computer Systems University of Toronto David Keppel EECS, UC Berkeley CS&amp;E, University of Washington Mark Brader SoftQuad Incorporated Toronto 1. Introduction This document is a modified version of a document from a committee formed at AT&amp;T&apos;s Indian Hill labs to establish a common set of coding standards and recommendations for the Indian Hill community. The scope of this work is C coding style. Good style should encourage consistent layout, improve portability, and reduce errors. This work does not cover functional organization, or general ..