Quantitative N-Glycan Profiling of Therapeutic Monoclonal Antibodies Performed by Middle-Up Level HILIC-HRMS Analysis

The identification and accurate quantitation of the various glycoforms contained in therapeutic monoclonal antibodies (mAbs) is one of the main analytical needs in the biopharmaceutical industry, and glycosylation represents a crucial critical quality attribute (CQA) that needs to be addressed. Currently, the reference method for performing such identification/quantitation consists of the release of the N-glycan moieties from the mAb, their labelling with a specific dye (e.g., 2-AB or RFMS) and their analysis by HILIC-FLD or HILIC-MS. In this contribution, the potential of a new cost- and time-effective analytical approach performed at the protein subunit level (middle-up) was investigated for quantitative purposes and compared with the reference methods. The robustness of the approach was first demonstrated by performing the relative quantification of the glycoforms related to a well characterized mAb, namely adalimumab. Then, the workflow was applied to various glyco-engineered mAb products (i.e., obinutuzumab, benralizumab and atezolizumab). Finally, the glycosylation pattern of infliximab (Remicade®) was assessed and compared to two of its commercially available biosimilars (Remsima® and Inflectra®). The middle-up analysis proved to provide accurate quantitation results and has the added potential to be used as multi-attribute monitoring method.

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LC-MS determination of glycosylation pattern on glycoproteins as critical quality attribute for biopharmaceuticals and potential markers for diseases

10.17760/d20195696 ◽

2015 ◽

Author(s):

Anna (Fan) Zhang

Keyword(s):

Quality Attribute ◽

Critical Quality Attribute ◽

Glycosylation Pattern

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Development of a chromatography-free method for high-throughput MS-based bioanalysis of therapeutic monoclonal antibodies

Bioanalysis ◽

10.4155/bio-2021-0021 ◽

2021 ◽

Author(s):

Zhengqi Zhang ◽

Yuetian Yan ◽

Shunhai Wang ◽

Ning Li

Keyword(s):

Monoclonal Antibodies ◽

High Throughput ◽

Solid Phase ◽

Proof Of Concept ◽

Direct Infusion ◽

Monitoring Method ◽

Parallel Reaction Monitoring ◽

Human Igg1 ◽

Surrogate Peptide ◽

Therapeutic Monoclonal Antibodies

Aim: Our objective was to test the feasibility of developing an LC-free, MS-based approach for high-throughput bioanalysis of humanized therapeutic monoclonal antibodies. Methodology: A universal tryptic peptide from human IgG1, IgG3 and IgG4 was selected as the surrogate peptide for quantitation. After tryptic digestion, the surrogate peptide was fractionated via solid-phase extraction before being subjected to direct infusion-based MS/MS analysis. A high-resolution, multiplexed (MSX = 2) parallel reaction monitoring method was developed for data acquisition. Results & conclusion: This proof-of-concept study demonstrated the feasibility of achieving high-throughput MS-based bioanalysis of monoclonal antibodies using an LC-free workflow with sensitivity comparable to conventional LC–MS/MS-based methods.

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Using Physiologically Based Pharmacokinetic (PBPK) Modeling as a Supportive Tool for Risk Assessment on the Critical Quality Attribute of a Generic Drug Product

10.26226/morressier.57d6b2bcd462b8028d88e03d ◽

2016 ◽

Author(s):

Fang Wu

Keyword(s):

Risk Assessment ◽

Generic Drug ◽

Drug Product ◽

Pbpk Modeling ◽

Quality Attribute ◽

Critical Quality Attribute ◽

Physiologically Based Pharmacokinetic ◽

Generic Drug Product ◽

Physiologically Based

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Design of Innovative Therapeutic Monoclonal Antibodies

Biotekhnologiya ◽

10.21519/0234-2758-2017-33-1-10-29 ◽

2017 ◽

pp. 10-29

Author(s):

A.V. Karabelskii ◽

◽

T.A. Nemankin ◽

A.B. Ulitin ◽

A.S. Vaganov ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Therapeutic Monoclonal Antibodies

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Therapeutic Monoclonal Antibodies in Clinical Practice against Cancer

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520620666200703191653 ◽

2020 ◽

Vol 20 (16) ◽

pp. 1895-1907

Author(s):

Navgeet Kaur ◽

Anju Goyal ◽

Rakesh K. Sindhu

Keyword(s):

Monoclonal Antibody ◽

Monoclonal Antibodies ◽

Clinical Practice ◽

Therapeutic Agent ◽

Hematologic Malignancies ◽

Immune Checkpoints ◽

Malignant Cells ◽

Main Target ◽

Therapeutic Monoclonal Antibodies ◽

Cancerous Cells

The importance of monoclonal antibodies in oncology has increased drastically following the discovery of Milstein and Kohler. Since the first approval of the monoclonal antibody, i.e. Rituximab in 1997 by the FDA, there was a decline in further applications but this number has significantly increased over the last three decades for various therapeutic applications due to the lesser side effects in comparison to the traditional chemotherapy methods. Presently, numerous monoclonal antibodies have been approved and many are in queue for approval as a strong therapeutic agent for treating hematologic malignancies and solid tumors. The main target checkpoints for the monoclonal antibodies against cancer cells include EGFR, VEGF, CD and tyrosine kinase which are overexpressed in malignant cells. Other immune checkpoints like CTLA-4, PD-1 and PD-1 receptors targeted by the recently developed antibodies increase the capability of the immune system in destroying the cancerous cells. Here, in this review, the mechanism of action, uses and target points of the approved mAbs against cancer have been summarized.

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COVID-19: Unmasking Emerging SARS-CoV-2 Variants, Vaccines and Therapeutic Strategies

Biomolecules ◽

10.3390/biom11070993 ◽

2021 ◽

Vol 11 (7) ◽

pp. 993

Author(s):

Renuka Raman ◽

Krishna J. Patel ◽

Kishu Ranjan

Keyword(s):

Immune Response ◽

Monoclonal Antibodies ◽

Severe Acute Respiratory Syndrome ◽

Small Molecules ◽

Viral Vector ◽

Therapeutic Strategies ◽

Convincing Evidence ◽

Plasma Therapy ◽

Therapeutic Monoclonal Antibodies ◽

Increased Susceptibility

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the etiological agent of the coronavirus disease 2019 (COVID-19) pandemic, which has been a topic of major concern for global human health. The challenge to restrain the COVID-19 pandemic is further compounded by the emergence of several SARS-CoV-2 variants viz. B.1.1.7 (Alpha), B.1.351 (Beta), P1 (Gamma) and B.1.617.2 (Delta), which show increased transmissibility and resistance towards vaccines and therapies. Importantly, there is convincing evidence of increased susceptibility to SARS-CoV-2 infection among individuals with dysregulated immune response and comorbidities. Herein, we provide a comprehensive perspective regarding vulnerability of SARS-CoV-2 infection in patients with underlying medical comorbidities. We discuss ongoing vaccine (mRNA, protein-based, viral vector-based, etc.) and therapeutic (monoclonal antibodies, small molecules, plasma therapy, etc.) modalities designed to curb the COVID-19 pandemic. We also discuss in detail, the challenges posed by different SARS-CoV-2 variants of concern (VOC) identified across the globe and their effects on therapeutic and prophylactic interventions.

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