Discovery and development of biopharmaceuticals: current issues

doi:10.1016/j.copbio.2009.10.012

Current Opinion in Biotechnology

Volume 20, Issue 6, December 2009, Pages 668-672

https://doi.org/10.1016/j.copbio.2009.10.012 Get rights and content

The establishment of recombinant DNA technologies in the past several decades has made it possible to develop a wide variety of therapeutic proteins to address a corresponding variety of unmet medical needs. Therapeutic proteins have had a major impact on health care, and are likely to grow in importance in the future. As the field of biotherapeutics evolves, there are many current and future challenges to be met. A highly competitive environment has emerged, in many cases with multiple products designed to modulate the same therapeutic target or pathway. The competition is resulting in more effective therapeutics as the need to differentiate among similar products becomes crucial. The costs and timelines for development of therapeutic proteins are increasing dramatically, leading to the tendency to focus primarily on those molecules with the greatest chance for success in late-stage clinical programs. As the field of therapeutic proteins continues to mature, the challenge of follow-on biologics needs to be addressed with the establishment of regulatory guidelines appropriate for the complexity of biotherapeutics.

Section snippets

Introduction—impact of therapeutic proteins on health care

Therapeutic proteins have been around for nearly 115 years, since the first development of commercial ‘Blutserumtherapie’ (serum therapy) by Behring and Kitasato. Behring's diphtheria serum therapy was first tested clinically in 1891 and by 1894, Faberwerke Hoechst launched the first therapeutic ‘protein’, anti-diphtheria serum (e.g., crude polyclonal antibody preparation), to combat a serious diphtheria epidemic in Europe [1^••]. One of the next advances in therapeutic proteins was the

Challenges for therapeutic proteins

There are many current and future challenges for biologics, ranging from spiraling development costs, to increased competition for targets and therapeutic indications, to the greater need for differentiation, to immunogenicity issues, to increasing competition from follow-on biologics, and the pressure on pricing from third party payors [1••, 5]. In this introductory paper, we will briefly touch on three main challenges: competition including the increased need for differentiation,

References and recommended reading

Papers of particular interest, published within the annual period of the review, have been highlighted as:

• of special interest
•• of outstanding interest

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Cited by (73)

Optimizing zinc-HisTag coordination remote loading of proteins in PLGA microspheres
2022, International Journal of Pharmaceutics
Citation Excerpt :
This process can expose the protein molecules to stressors including micronization, organic/aqueous interfaces, air/water interfaces, high shear stress, organic solvents, and high temperatures (Ando et al., 1999; Cohen and Bernstein, 1996; Giovagnoli et al., 2004; Han et al., 2016; Reinhold et al., 2012; Schwendeman, 2002; Schwendeman et al., 1996; Wu and Jin, 2008). Another issue that is particularly challenging during drug discovery and early development is that the standard batch formulation process requires large amounts of the drug molecule, which can be cost-prohibitive or simply infeasible at these stages (Murray et al., 1984; Strohl and Knight, 2009). To avoid these stressors and reduce the mass of protein required during formulation, aqueous remote loading and self-healing encapsulation has been developed with our group being pioneers in the space (Reinhold et al., 2012; Schwendeman, 2005; Schwendeman et al., 2011; Shah and Schwendeman, 2014; Wang et al., 2002).
Metal-HisTag coordination remote loading (MHCRL) of proteins in PLGA microspheres was previously developed to provide a useful tool for discovery and preclinical development of controlled release protein formulations. Here we describe optimization of MHCRL, including (1) reducing thermal stress, (2) decreasing the complexity and duration of the procedure, (3) increasing loading capacity, (4) increasing the penetration depth of protein, and (5) improving the release profile. Directly encapsulating ZnCO₃ as a Zn²⁺ source for HisTag coordination, rather than remotely loading Zn²⁺, increased the Zn content ∼6-fold. Microspheres with directly encapsulated ZnCO₃ more deeply encapsulated green fluorescent protein and more efficiently encapsulated human serum albumin at protein loading solutions concentrations ≥100 μg/mL than remotely loaded Zn²⁺ microspheres. Tributyl acetylcitrate plasticized microspheres in terms of decreasing T_g, but led to a decrease in protein encapsulation efficiency. As such, the plasticizer was not deemed useful. The loading/healing cycles were reduced in time and temperature from 48 h/42 h at 43 °C to 2 h/6h at 37 °C while maintaining strong encapsulation efficiency, resulting in significantly improved protein stability. Immunoreactive protein was slowly released for months following a modest burst release. The improved microspheres and shorter, low-temperature encapsulation could be a valuable asset to drug discovery scientists interested in controlled release of delicate and/or costly biologic candidates.
Recombinant Elabela-Fc fusion protein has extended plasma half-life and mitigates post-infarct heart dysfunction in rats
2020, International Journal of Cardiology
Combination of phage and Gram-positive bacterial display of human antibody repertoires enables isolation of functional high affinity binders
2018, New Biotechnology
Surface display couples genotype with a surface exposed phenotype and thereby allows screening of gene-encoded protein libraries for desired characteristics. Of the various display systems available, phage display is by far the most popular, mainly thanks to its ability to harbour large size libraries. Here, we describe the first use of a Gram-positive bacterial host for display of a library of human antibody genes which, when combined with phage display, provides ease of use for screening, sorting and ranking by flow cytometry. We demonstrate the utility of this method by identifying low nanomolar affinity scFv fragments towards human epidermal growth factor receptor 2 (HER2). The ranking and performance of the scFv isolated by flow sorting in surface-immobilised form was retained when expressed as soluble scFv and analysed by biolayer interferometry, as well as after expression as full-length antibodies in mammalian cells. We also demonstrate the possibility of using Gram-positive bacterial display to directly improve the affinity of the identified binders via an affinity maturation step using random mutagenesis and flow sorting. This combined approach has the potential for a more complete scan of the antibody repertoire and for affinity maturation of human antibody formats.
Increased cryosurvival of osteosarcoma cells using an amphipathic pH-responsive polymer for trehalose uptake
2016, Cryobiology
Amphipathic pH-responsive polymers have shown to increase the permeability of cell membranes to trehalose hence improving the cryopreservation of mammalian cells. However, the trafficking of both the polymer and trehalose across the cell membrane has not yet been thoroughly analysed. The objective of this study was to investigate the effect on cryopreservation of the trafficking of the disaccharide trehalose along PP-50, an amphipathic polymer, through an osteosarcoma cell line (SAOS-2). Confocal microscopy analysis confirmed the presence of intracellular labelled trehalose only when incubated in the presence of PP-50. Further analysis confirmed that both trehalose and PP-50 localised in the cytoplasm, accumulated mainly in the perinuclear area. Quantitative analysis of the colocalisation between trehalose and PP-50 showed Pearson and Manders coefficients of 0.862 ± 0.008 and 0.766 ± 0.033, respectively, suggesting a high degree of intracellular colocalisation between these molecules. Cryopreserved cells pre-incubated with trehalose and PP-50 showed increased cryosurvival when compared with cells pre-incubated in the absence of the polymer. PP-50 showed to be directly involved in the uptake of trehalose, a critical characteristic towards use in cryopreservation and biomedical applications.
Potential of hydrophilic interaction chromatography for the analytical characterization of protein biopharmaceuticals
2016, Journal of Chromatography A
A new stationary phase based on wide-pore hybrid silica bonded with amide ligand has been used to explore the utility of HILIC for the analytical characterization of protein biopharmaceuticals. Various, highly-relevant samples were tested, including different insulins, interferon α-2b and trastuzumab. This work shows that HILIC can be successfully employed for the analysis of therapeutic proteins and mAbs, using mobile phase compositions comprised of between 65 and 80% ACN and 0.1% TFA. In terms of elution order and selectivity, these HILIC separations have proven to be highly orthogonal to RPLC, while the kinetic performance remains comparable. In the case of characterizing trastuzumab, HILIC was uniquely able to resolve several important glycoforms at the middle-up level of analysis (fragments of 25–100 kDa). Such a separation of glycoforms has been elusive by other separation mechanisms, such as RPLC and IEX. Besides showing orthogonality to RPLC and improved separations of glycoforms, HILIC offers several additional benefits for biopharmaceutical characterization: i) an inherent compatibility with MS, ii) a reduced requirement for very high mobile phase temperatures that are otherwise needed in RPLC to limit undesirably strong adsorption to the surface of the stationary phase, and iii) the possibility to couple several columns in series to improve resolving power, thanks to comparatively low mobile phase viscosity.
Benefits of using heterologous polyclonal antibodies and potential applications to new and undertreated infectious pathogens
2016, Vaccine
Passive immunotherapy using polyclonal antibodies (immunoglobulins) has been used for over a century in the treatment and post-exposure prophylaxis of various infections and toxins. Heterologous polyclonal antibodies are obtained from animals hyperimmunised with a pathogen or toxin.
The aims of this review are to examine the history of animal polyclonal antibody therapy use, their development into safe and effective products and the potential application to humans for emerging and neglected infectious diseases.
A literature search of OVID Medline and OVID Embase databases was undertaken to identify articles on the safety, efficacy and ongoing development of polyclonal antibodies. The search contained database-specific MeSH and EMTREE terms in combination with pertinent text-words: polyclonal antibodies and rare/neglected diseases, antivenins, immunoglobulins, serum sickness, anaphylaxis, drug safety, post marketing surveillance, rabies, human influenza, Dengue, West Nile, Nipah, Hendra, Marburg, MERS, Hemorrhagic Fever Virus, and Crimean-Congo. No language limits were applied. The final search was completed on 20.06.2015. Of 1960 articles, title searches excluded many irrelevant articles, yielding 303 articles read in full. Of these, 179 are referenced in this study.
Serum therapy was first used in the 1890s against diphtheria. Early preparation techniques yielded products contaminated with reactogenic animal proteins. The introduction of enzymatic digestion, and purification techniques substantially improved their safety profile. The removal of the Fc fragment of antibodies further reduces hypersensitivity reactions. Clinical studies have demonstrated the efficacy of polyclonal antibodies against various infections, toxins and venoms. Products are being developed against infections for which prophylactic and therapeutic options are currently limited, such as avian influenza, Ebola and other zoonotic viruses.
Polyclonal antibodies have been successfully applied to rabies, envenomation and intoxication. Polyclonal production provides an exciting opportunity to revolutionise the prognosis of both longstanding neglected tropical diseases as well as emerging infectious threats to humans.

View all citing articles on Scopus

Dr. William R. Strohl received his PhD in microbiology from Louisiana State University, and worked as a guest researcher at the GBF in Braunschweig, Germany. From 1980 to 1997, he rose from assistant to full professor in the Department of Microbiology and the Program of Biochemistry at The Ohio State University, Columbus, OH. There he pursued the molecular biology and biochemistry of polyketide biosynthesis pathways, particularly doxorubicin, in actinomycetes, and the physiology of E. coli in computer-controlled high cell density fermentations. Dr. Strohl moved to Merck in 1997 to head up Natural Products Microbiology, which he did for 4 years before being asked to start a new department in the field of recombinant monoclonal antibodies. From 2001 to 2008, Dr. Strohl was a leader in Merck's efforts to develop therapeutic monoclonal antibodies, as well as in-licensing of therapeutic targets and technologies associated with monoclonal antibodies. As part of this effort, Dr. Strohl was the scientific leader in the acquisition and integration of Abmaxis and GlycoFi into the Merck Biologics organization. In April 2008, Dr. Strohl was named leader of Antibody Drug Discovery at Centocor. With evolution of biologics at Johnson & Johnson to serve additional therapeutic areas beyond Immunology and Oncology, Dr. Strohl was recently named as VP and Head, Biologics Research, in the newly formed J&J Biotechnology Center of Excellence. Dr. Strohl has over 100 publications and several patents, and has edited two books.

Dr. Knight received his PhD in molecular biology from the University of California, Berkeley, and completed a postdoctoral fellowship at Stanford University School of Medicine. He joined Centocor in 1985, rising to the position of Vice President, Biopharmaceutical Research. Dr. Knight played key roles in the discovery and development of Centocor's marketed biotherapeutics including ReoPro™, Remicade™. Simponi™, and Stelara™. He is the author of numerous publications and holds more than 50 issued US patents. Centocor became part of Johnson and Johnson in 1999, and Dr. Knight is currently a Distinguished Research Fellow in the J&J Biotechnology Center of Excellence.

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Editorial overviewDiscovery and development of biopharmaceuticals: current issues

Section snippets

Introduction—impact of therapeutic proteins on health care

Challenges for therapeutic proteins

References and recommended reading

J Clin Oncol

Hematol Am Soc Hematol Educ Program

Curr Opin Immunol

J Immunol

Protein therapeutics: a summary and pharmacological classification

Nat Rev Drug Discov

Protein therapeutics—lessons learned and a view of the future

Expert Opin Biol Ther

Brand biologics grab 12 years’ exclusivity, for now

Nat Biotechnol

Biosimilars: it's not as simple as cost alone

J Clin Pharm Ther

Editorial overview
Discovery and development of biopharmaceuticals: current issues