Challenges and Emerging Technologies in Biomanufacturing of Monoclonal Antibodies (mAbs)

McDonnell, Susan; Principe, Raymon Floyd; Zamprognio, Maycou Soares; Whelan, Jessica

doi:10.5772/intechopen.108565

Cited by 8 publications

(4 citation statements)

References 78 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…13 Commercial development and manufacturing of mAb biotherapeutics need to meet the strict standards of regulatory authorities such as the Food and Drug Administration (FDA) and the European Medicines Agency (EMA) before entering the market. 14,15 The standards set by the FDA and EMA are based on the Quality by Design (QbD) approach, used to assess the impact of various variables on the critical quality attributes (CQA), such as aggregates and glycosylation of a drug product. [15][16][17] Understanding how process parameters can affect CQAs is an important step the FDA has been investigating for the past couple of years.…”

Section: Introductionmentioning

confidence: 99%

Hybrid mass spectrometry applied across the production of antibody biotherapeutics.

Christofi,

O'Hanlon,

Curtis

et al. 2024

Preprint

View full text Add to dashboard Cite

Post expression from the host cell, a bio-therapeutic will be subjected to several downstream processing steps prior to final formulation. Analysis of the product under various buffer conditions at each stage is desirable to show that the integrity of the sample is maintained and to monitor any potential product loss. In this study we examine how varying buffer conditions during downstream processing impact the structural integrity of a biopharmaceutical product, with a specific focus on the model IgG1 antibody, mAb4, provided by FUJIFILM Diosynth Biotechnologies. Flexibility, stability, aggregation propensity, and bulk properties were evaluated across the buffers of four sample points, including perfusion media, purification stages, and formulation buffer. Comparisons with Herceptin, an extensively studied IgG1 antibody, were conducted in a mass spectrometry-compatible buffer. Although mAb4 and Herceptin present with similar charge state distributions (CSD) in native mass spectrometry (MS) experiments, they exhibit distinct unfolding patterns during the activated ion mobility-mass spectrometry (aIM-MS) and differential scanning fluorimetry (DSF). The greater structural stability and aggregation onset temperature (Tagg) observed in Herceptin, are primarily attributed to the heavier glycosylation and kappa-class light chains present in Herceptin compared to the lambda-class light chains in mAb4. Hydrogen deuterium exchange mass spectrometry (HDX-MS) revealed that while mAb4 experiences substantial structural changes during purification, characterized by high flexibility, a low melting temperature (Tm), and prevalent repulsive protein-protein interactions, it eventually transitions to a highly compact and stable structure in buffers with higher salt concentrations and during formulation. Notably, in the formulated environment, the third constant domain (CH3) of the heavy chain retains flexibility and is identified as a region of interest for aggregation formation. Here we showcase the integration of MS and orthogonal techniques to attain comprehensive information which can be utilized early in the development stage to aid in decision-making regarding targeted mutations or to guide the design space of bioprocesses and formulation choices.

show abstract

Section: Introductionmentioning

confidence: 99%

Hybrid mass spectrometry applied across the production of antibody biotherapeutics.

Christofi,

O'Hanlon,

Curtis

et al. 2024

Preprint

View full text Add to dashboard Cite

show abstract

“…The optimization of therapeutic antibodies by conventional experiments is time-consuming and laborious [ 12 , 13 ]. For example, low-throughput screening of full-length antibodies based on mammalian systems expressed in cells usually results in a problem of affinity ceiling of an antibody [ 14 ], while in vitro methods for affinity maturation are more sensible but inefficient [ 12 , 15 ].…”

Section: Introductionmentioning

confidence: 99%

AttABseq: an attention-based deep learning prediction method for antigen–antibody binding affinity changes based on protein sequences

Jin,

Ye,

Wang

et al. 2024

Briefings in Bioinformatics

View full text Add to dashboard Cite

The optimization of therapeutic antibodies through traditional techniques, such as candidate screening via hybridoma or phage display, is resource-intensive and time-consuming. In recent years, computational and artificial intelligence-based methods have been actively developed to accelerate and improve the development of therapeutic antibodies. In this study, we developed an end-to-end sequence-based deep learning model, termed AttABseq, for the predictions of the antigen–antibody binding affinity changes connected with antibody mutations. AttABseq is a highly efficient and generic attention-based model by utilizing diverse antigen–antibody complex sequences as the input to predict the binding affinity changes of residue mutations. The assessment on the three benchmark datasets illustrates that AttABseq is 120% more accurate than other sequence-based models in terms of the Pearson correlation coefficient between the predicted and experimental binding affinity changes. Moreover, AttABseq also either outperforms or competes favorably with the structure-based approaches. Furthermore, AttABseq consistently demonstrates robust predictive capabilities across a diverse array of conditions, underscoring its remarkable capacity for generalization across a wide spectrum of antigen-antibody complexes. It imposes no constraints on the quantity of altered residues, rendering it particularly applicable in scenarios where crystallographic structures remain unavailable. The attention-based interpretability analysis indicates that the causal effects of point mutations on antibody–antigen binding affinity changes can be visualized at the residue level, which might assist automated antibody sequence optimization. We believe that AttABseq provides a fiercely competitive answer to therapeutic antibody optimization.

show abstract

“…Similarly, the FDA has authorized two antibody cocktails, tixagevimab/cilgavimab and casirivimab/ imdevimab, for clinical use in COVID -19 treatment (11, 14). Nonetheless, mAb cocktail approaches have limitations, including heightened manufacturing costs, time-consuming processes, and high-dose infusions for patients (25,26). Thus, bsAbs emerge as a promising strategy to confront ever-evolving SARS-CoV-2 variants.…”

Section: Introductionmentioning

confidence: 99%

A novel bispecific antibody dual-targeting approach for enhanced neutralization against fast-evolving SARS-CoV-2 variants

Kim,

Heo

et al. 2023

Front. Immunol.

View full text Add to dashboard Cite

IntroductionThe emergence of new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants has caused unprecedented health and socioeconomic crises, necessitating the immediate development of highly effective neutralizing antibodies. Despite recent advancements in anti-SARS-CoV-2 receptor-binding domain (RBD)-specific monoclonal antibodies (mAbs) derived from convalescent patient samples, their efficacy against emerging variants has been limited. In this study, we present a novel dual-targeting strategy using bispecific antibodies (bsAbs) that specifically recognize both the SARS-CoV-2 RBD and fusion peptide (FP), crucial domains for viral attachment to the host cell membrane and fusion in SARS-CoV-2 infection. MethodsUsing phage display technology, we rapidly isolated FP-specific mAbs from an established human recombinant antibody library, identifying K107.1 with a nanomolar affinity for SARS-CoV-2 FP. Furthermore, we generated K203.A, a new bsAb built in immunoglobulin G4-(single-chain variable fragment)2 forms and demonstrating a high manufacturing yield and nanomolar affinity to both the RBD and FP, by fusing K102.1, our previously reported RBD-specific mAb, with K107.1. ResultsOur comprehensive in vitro functional analyses revealed that the K203.A bsAb significantly outperformed the parental RBD-specific mAb in terms of neutralization efficacy against SARS-CoV-2 variants. Furthermore, intravenous monotherapy with K203.A demonstrated potent in vivo neutralizing activity without significant in vivo toxicity in a mouse model infected with a SARS-CoV-2 variant. ConclusionThese findings present a novel bsAb dual-targeting strategy, directed at SARS-CoV-2 RBD and FP, as an effective approach for rapid development and management against continuously evolving SARS-CoV-2 variants.

show abstract

Challenges and Emerging Technologies in Biomanufacturing of Monoclonal Antibodies (mAbs)

Cited by 8 publications

References 78 publications

Hybrid mass spectrometry applied across the production of antibody biotherapeutics.

Hybrid mass spectrometry applied across the production of antibody biotherapeutics.

AttABseq: an attention-based deep learning prediction method for antigen–antibody binding affinity changes based on protein sequences

A novel bispecific antibody dual-targeting approach for enhanced neutralization against fast-evolving SARS-CoV-2 variants

Contact Info

Product

Resources

About