Excipients Used in Biotechnology Products

Y., Eva

doi:10.1002/9781118992432.ch4

Cited by 9 publications

(4 citation statements)

References 141 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This idea is bolstered by examining the additives that protect against freeze-dry stress but not desiccation stress: the reducing sugars, dextran, maltose, and glucose. This lack of protection may arise from covalent modification by these sugars (glycation, a known problem in formulation) as their concentration increases during desiccation. Further support for the suggestion that different mechanisms operate for freeze-thaw- and desiccation-induced inactivation comes from the observation that lysozyme denatures LDH in concentrated solutions, but dilute lysozyme solutions can fully protect against freeze–thaw inactivation …”

Section: Analysis and Conclusionmentioning

confidence: 99%

Protecting Enzymes from Stress-Induced Inactivation

Piszkiewicz

Pielak

2019

Biochemistry

View full text Add to dashboard Cite

The pharmaceutical and chemical industries depend on additives to protect enzymes and other proteins against stresses that accompany their manufacture, transport, and storage. Common stresses include vacuum-drying, freeze-thawing, and freeze-drying. The additives include sugars, compatible osmolytes, amino acids, synthetic polymers, and both globular and disordered proteins. Scores of studies have been published on protection, but the data have never been analyzed systematically. To spur efforts to understand the sources of protection and ultimately develop more effective formulations, we review ideas about the mechanisms of protection, survey the literature searching for patterns of protection, and then compare the ideas to the data.

show abstract

Section: Analysis and Conclusionmentioning

confidence: 99%

Protecting Enzymes from Stress-Induced Inactivation

Piszkiewicz

Pielak

2019

Biochemistry

View full text Add to dashboard Cite

show abstract

“…Proteins are only marginally stable and prone to physical degradation. Thus, an excipient was added to the r28M to enhance its stability by suppressing aggregation, the primary pathway of protein physical degradation, surface adsorption or by maintaining physiological osmolarity [ 42 ]. The positive effects of excipients have often been described [ 43 ] and the European Medicines Agency (EMA) provides guidance on excipients for application for marketing authorization of medicinal products.…”

Section: Discussionmentioning

confidence: 99%

Large-Scale Purification of r28M: A Bispecific scFv Antibody Targeting Human Melanoma Produced in Transgenic Cattle

et al. 2015

View full text Add to dashboard Cite

Background30 years ago, the potential of bispecific antibodies to engage cytotoxic T cells for the lysis of cancer cells was discovered. Today a variety of bispecific antibodies against diverse cell surface structures have been developed, the majority of them produced in mammalian cell culture systems. Beside the r28M, described here, no such bispecific antibody is known to be expressed by transgenic livestock, although various biologicals for medical needs are already harvested—mostly from the milk—of these transgenics. In this study we investigated the large-scale purification and biological activity of the bispecific antibody r28M, expressed in the blood of transgenic cattle. This tandem single-chain variable fragment antibody is designed to target human CD28 and the melanoma/glioblastoma-associated cell surface chondroitin sulfate proteoglycan 4 (CSPG4).ResultsWith the described optimized purification protocol an average yield of 30 mg enriched r28M fraction out of 2 liters bovine plasma could be obtained. Separation of this enriched fraction by size exclusion chromatography into monomers, dimers and aggregates and further testing regarding the biological activity revealed the monomer fraction as being the most appropriate one to continue working with. The detailed characterization of the antibody’s activity confirmed its high specificity to induce the killing of CSPG4 positive cells. In addition, first insights into tumor cell death pathways mediated by r28M-activated peripheral blood mononuclear cells were gained. In consideration of possible applications in vivo we also tested the effect of the addition of different excipients to r28M.ConclusionSumming up, we managed to purify monomeric r28M from bovine plasma in a large-scale preparation and could prove that its biological activity is unaffected and still highly specific and thus, might be applicable for the treatment of melanoma.

show abstract

“…Liquid formulations currently represent the most common administration route for monoclonal antibodies . These formulations contain a variety of excipients to maintain pH and tonicity and to increase protein stability and preservation. − Typical categories of excipients include buffering agents, tonicity modifiers, thermal stabilizers, surfactants, and amino acids. − This strategy offers many degrees of freedom since in principle an infinite number of different excipients and their combinations can be selected to simultaneously optimize multiple properties of a protein. The introduction of novel excipients, however, is largely prevented by regulatory considerations and corresponding increases in the approval timelines. ,, As a consequence, excipients are typically selected from the approved list of molecules documented under the FDA’s inactive substance database, which are generally regarded as safe (GRAS) …”

Section: Introductionmentioning

confidence: 99%

Design of Biopharmaceutical Formulations Accelerated by Machine Learning

et al. 2021

View full text Add to dashboard Cite

In addition to activity, successful biological drugs must exhibit a series of suitable developability properties, which depend on both protein sequence and buffer composition. In the context of this high-dimensional optimization problem, advanced algorithms from the domain of machine learning are highly beneficial in complementing analytical screening and rational design. Here, we propose a Bayesian optimization algorithm to accelerate the design of biopharmaceutical formulations. We demonstrate the power of this approach by identifying the formulation that optimizes the thermal stability of three tandem single-chain Fv variants within 25 experiments, a number which is less than one-third of the experiments that would be required by a classical DoE method and several orders of magnitude smaller compared to detailed experimental analysis of full combinatorial space. We further show the advantage of this method over conventional approaches to efficiently transfer historical information as prior knowledge for the development of new biologics or when new buffer agents are available. Moreover, we highlight the benefit of our technique in engineering multiple biophysical properties by simultaneously optimizing both thermal and interface stabilities. This optimization minimizes the amount of surfactant in the formulation, which is important to decrease the risks associated with corresponding degradation processes. Overall, this method can provide high speed of converging to optimal conditions, the ability to transfer prior knowledge, and the identification of new nonlinear combinations of excipients. We envision that these features can lead to a considerable acceleration in formulation design and to parallelization of operations during drug development.

show abstract

Excipients Used in Biotechnology Products

Cited by 9 publications

References 141 publications

Protecting Enzymes from Stress-Induced Inactivation

Protecting Enzymes from Stress-Induced Inactivation

Large-Scale Purification of r28M: A Bispecific scFv Antibody Targeting Human Melanoma Produced in Transgenic Cattle

Design of Biopharmaceutical Formulations Accelerated by Machine Learning

Contact Info

Product

Resources

About