Native-State Solubility and Transfer Free Energy as Predictive Tools for Selecting Excipients to Include in Protein Formulation Development Studies

Banks, Douglas D.; Latypov, Ramil F.; Ketchem, Randal R.; Woodard, Jon; Scavezze, Joanna L.; Siska, Christine C.; Razinkov, Vladimir I.

doi:10.1002/jps.23219

Cited by 63 publications

(62 citation statements)

References 51 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…[17][18][19][20][21] Lin et al recently demonstrated that limited information is provided by far-UV circular dichroism and Fourier transform infrared spectroscopy compared with SEC and IEX read outs in comparability studies with degraded mAb samples. 22 In an effort to determine the ability of other biophysical assays to predict product stability at different conditions, we retrospectively compared the initial molecular properties of 9 mAbs in their lead (i.e., optimized) formulations with their stability behavior.…”

Section: Discussionmentioning

confidence: 99%

A comparison of biophysical characterization techniques in predicting monoclonal antibody stability

Thiagarajan

Semple

James

et al. 2016

mAbs

103

View full text Add to dashboard Cite

With the rapid growth of biopharmaceutical product development, knowledge of therapeutic protein stability has become increasingly important. We evaluated assays that measure solution-mediated interactions and key molecular characteristics of 9 formulated monoclonal antibody (mAb) therapeutics, to predict their stability behavior. Colloidal interactions, self-association propensity and conformational stability were measured using effective surface charge via zeta potential, diffusion interaction parameter (k D ) and differential scanning calorimetry (DSC), respectively. The molecular features of all 9 mAbs were compared to their stability at accelerated (25 C and 40 C) and long-term storage conditions (2-8 C) as measured by size exclusion chromatography. At accelerated storage conditions, the majority of the mAbs in this study degraded via fragmentation rather than aggregation. Our results show that colloidal stability, self-association propensity and conformational characteristics (exposed tryptophan) provide reasonable prediction of accelerated stability, with limited predictive value at 2-8 C stability. While no correlations to stability behavior were observed with onset-of-melting temperatures or domain unfolding temperatures, by DSC, melting of the Fab domain with the C H 2 domain suggests lower stability at stressed conditions. The relevance of identifying appropriate biophysical assays based on the primary degradation pathways is discussed.

show abstract

Section: Discussionmentioning

confidence: 99%

A comparison of biophysical characterization techniques in predicting monoclonal antibody stability

Thiagarajan

Semple

James

et al. 2016

mAbs

103

View full text Add to dashboard Cite

show abstract

“…These may become exposed transiently via local or partial unfolding (structural distortion) of an initially folded monomer, or if one first forms small, reversibly folded oligomers or “clusters” [18]. Under conditions in which the folded state is favored over the unfolded states, monomers can initially self-associate reversibly (Figure 1, left) either as folded or partially unfolded species.…”

Section: Mechanisms Dictate Formation Rates and Key Characteristics Omentioning

confidence: 99%

“…The former is supported by indirect assessments in which monomer conformational stability explains the temperature dependence of aggregation quantitatively or semi-quantitiatively [21–24]. The latter is supported by indirect assessments such as correlations of aggregation rates with the solubility (or chemical potential) of folded monomer [18]. …”

Section: Mechanisms Dictate Formation Rates and Key Characteristics Omentioning

confidence: 99%

Therapeutic protein aggregation: mechanisms, design, and control

Roberts

2014

Trends in Biotechnology

356

309

View full text Add to dashboard Cite

While it is well known that proteins are only marginally stable in their folded states, it is often less well appreciated that most proteins are inherently aggregation-prone in their unfolded or partially unfolded states, and the resulting aggregates can be extremely stable and long-lived. For therapeutic proteins, aggregates are a significant risk factor for deleterious immune responses in patients, and can form via a variety of mechanisms. Controlling aggregation using a mechanistic approach may allow improved design of therapeutic protein stability, as a complement to existing design strategies that target desired protein structures and function. Recent results highlight the importance of balancing protein environment with the inherent aggregation propensities of polypeptide chains.

show abstract

“…Nucleation is usually a kinetically limited process that can require extremely long time scales (from minutes to years) when compared to typical timescales for protein folding (~ μs to s) under native-favoring conditions where most proteins are stored and delivered. In many cases, partial unfolding of protein monomers appears to be the first step, followed by slow association of two or more chains to create the nuclei [37], although at high protein concentration it has been inferred that association occurs before unfolding [38,39], and in some cases unfolding is clearly rate-limiting. If growth occurs by monomer addition, then the (partially) unfolded monomers can add more easily to the existing aggregates [34,35,40].…”

Section: Why and How Do Proteins Aggregate?mentioning

confidence: 99%

Protein aggregation and its impact on product quality

Roberts

2014

Current Opinion in Biotechnology

243

175

View full text Add to dashboard Cite

Protein pharmaceutical products are typically active as folded monomers that are composed of one or more protein chains, such as the heavy and light chains in monoclonal antibodies that are a mainstay of current drug pipelines. There are numerous possible aggregated states for a given protein, some of which are potentially useful, while most of which are considered deleterious from the perspective of pharmaceutical product quality and performance. This review provides an overview of how and why different aggregated states of proteins occur, how this potentially impacts product quality and performance, fundamental approaches to control aggregate formation, and the practical approaches that are currently used in the pharmaceutical industry.

show abstract

Native-State Solubility and Transfer Free Energy as Predictive Tools for Selecting Excipients to Include in Protein Formulation Development Studies

Cited by 63 publications

References 51 publications

A comparison of biophysical characterization techniques in predicting monoclonal antibody stability

A comparison of biophysical characterization techniques in predicting monoclonal antibody stability

Therapeutic protein aggregation: mechanisms, design, and control

Protein aggregation and its impact on product quality

Contact Info

Product

Resources

About