Circular Dichroism Spectroscopy as a Tool for Monitoring Aggregation in Monoclonal Antibody Therapeutics

Joshi, Varsha; Shivach, Tarun; Yadav, Nitin; Rathore, Anurag S.

doi:10.1021/ac503140j

Cited by 106 publications

(78 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…At higher temperature, as the time progresses, the MRE values are continuously decreasing and this signifies that there is a conformational change in the protein. This also correlates with higher order aggregation observed at higher temperatures (25).…”

Section: Effect Of Temperaturesupporting

confidence: 73%

See 1 more Smart Citation

Aggregation Kinetics for IgG1-Based Monoclonal Antibody Therapeutics

Singla

Bansal

Joshi

et al. 2016

AAPS J

Self Cite

View full text Add to dashboard Cite

Abstract. Monoclonal antibodies (mAbs) as a class of therapeutic molecules are finding an increasing demand in the biotechnology industry for the treatment of diseases like cancer and multiple sclerosis. A key challenge associated to successful commercialization of mAbs is that from the various physical and chemical instabilities that are inherent to these molecules. Out of all probable instabilities, aggregation of mAbs has been a major problem that has been associated with a change in the protein structure and is a hurdle in various upstream and downstream processes. It can stimulate immune response causing protein misfolding having deleterious and harmful effects inside a cell. Also, the extra cost incurred to remove aggregated mAbs from the rest of the batch is huge. Size exclusion chromatography (SEC) is a major technique for characterizing aggregation in mAbs where change in the aggregates' size over time is estimated. The current project is an attempt to understand the rate and mechanism of formation of higher order oligomers when subjected to different environmental conditions such as buffer type, temperature, pH, and salt concentration. The results will be useful in avoiding the product exposure to conditions that can induce aggregation during upstream, downstream, and storage process. Extended Lumry-Eyring model (ELE), Lumry-Eyring Native Polymerization model (LENP), and Finke-Watzky model (F-W) have been employed in this work to fit the aggregation experimental data and results are compared to find the best fit model for mAb aggregation to connect the theoretical dots with the reality.

show abstract

Section: Effect Of Temperaturesupporting

confidence: 73%

“…Sample concentration was kept at 0.2 mg/ml and wavelength in the range of 200-250 nm was used to obtain spectra (25). For spectral measurements, quartz cuvette (1 mm path length) was used at 20°C and an average of five scans was taken.…”

Section: Circular Dichroism (Cd)mentioning

confidence: 99%

Aggregation Kinetics for IgG1-Based Monoclonal Antibody Therapeutics

Singla

Bansal

Joshi

et al. 2016

AAPS J

Self Cite

View full text Add to dashboard Cite

show abstract

“…Further deconvolution of CD spectrum using Dichroweb revealed that commercial and reconstituted BSA samples contained 57%–61% α–helix structure, which corresponds with known literature values (27, 50). The degradation or aggregation of protein could cause intensity change of CD spectrum or peak position shift (51). In a forced degradation study of BSA by Estey et al, BSA incubated in acidic media showed a loss of 13.7% α–helix structure (27).…”

Section: Resultsmentioning

confidence: 99%

A Multiparticulate Delivery System for Potential Colonic Targeting Using Bovine Serum Albumin as a Model Protein

Jiang

Zhang

et al. 2017

Pharm Res

View full text Add to dashboard Cite

Purpose There are many important diseases whose treatment could be improved by delivering a therapeutic protein to the colon, for example, Clostridium difficile infection, ulcerative colitis and Crohn’s Disease. The goal of this project was to investigate the feasibility of colonic delivery of proteins using multiparticulate beads. Methods In this work, bovine serum albumin (BSA) was adopted as a model protein. BSA was spray layered onto beads, followed by coating of an enteric polymer EUDRAGIT® FS 30 D to develop a colonic delivery system. The secondary and tertiary structure change and aggregation of BSA during spray layering process was examined. The BSA layered beads were then challenged in an accelerated stability study using International Council for Harmonization (ICH) conditions. The in vitro release of BSA from enteric coated beads was examined using United States Pharmacopeia (USP) dissolution apparatus 1. Results No significant changes in the secondary and tertiary structure or aggregation profile of BSA were observed after the spray layering process. Degradation of BSA to different extents was detected after storing at 25 °C and 40 °C for 38 days. Enteric coated BSA beads were intact in acidic media while released BSA in pH 7.4 phosphate buffer. Conclusion We showed the feasibility of delivering proteins to colon in vitro using multiparticulate system.

show abstract

“…Circular dichroïsm was recently described as a powerful tool to detect aggregates formation in mAbs preparations 39 . The interest of this method is the evaluation of structural changes, comparing native and stressed proteins.…”

Section: Discussionmentioning

confidence: 99%

Effect of growth hormone and IgG aggregates on dendritic cells activation and T‐cells polarization

et al. 2016

View full text Add to dashboard Cite

Patients treated with therapeutic biological products (BP) frequently develop anti-drug antibodies (ADA) with potential neutralizing capacities leading to loss of clinical response or serious side effects. BP aggregates have been suggested to promote immunogenicity, thus enhancing ADA production. Dendritic cells (DC) are key effectors in T-cell and B-cell fates, and the subsequent generation of immunogenicity. The objective of this work was to determine if BP aggregates can participate to DC maturation and T-cell activation. We compared aggregates from three different proteins: human growth hormone (hGH), Rituximab, a chimeric anti-CD20 antibody and a serum-purified human IgG1. All three proteins underwent a stir stress, generating comparable populations of aggregated particles. Maturation of human monocyte-derived DC (moDC) upon exposure to native BPs or aggregates was evaluated in vitro. Results showed that hGH aggregates induced an increased expression of moDC co-stimulation markers, and augmented levels of IL-6, IL-8, IL-12p40, CCL2, CCL3, CCL4 and CXCL10. Both antibodies aggregates were also able to modify DC phenotype, but cytokine and chemokine productions were seen only with IL-6, IL-8, IL-12p40 and CXCL10. Aggregates-treated moDC enhanced allogenic T-cell proliferation and cytokines production, suggesting Th1 polarization with hGH, and mixed T-cell responses with antibodies aggregates. These results showed that BP aggregates provoked DC maturation, thus driving adaptive T-cell responses and polarization.

show abstract

Circular Dichroism Spectroscopy as a Tool for Monitoring Aggregation in Monoclonal Antibody Therapeutics

Cited by 106 publications

References 30 publications

Aggregation Kinetics for IgG1-Based Monoclonal Antibody Therapeutics

Aggregation Kinetics for IgG1-Based Monoclonal Antibody Therapeutics

A Multiparticulate Delivery System for Potential Colonic Targeting Using Bovine Serum Albumin as a Model Protein

Effect of growth hormone and IgG aggregates on dendritic cells activation and T‐cells polarization

Contact Info

Product

Resources

About