Methods to measure the binding of therapeutic monoclonal antibodies to the human Fc receptor FcγRIII (CD16) using real time kinetic analysis and flow cytometry

Aim: Evaluation of suitable pharmacokinetic properties is critical for successful development of IgG-based biotherapeutics. The prolonged half-lives of IgGs depend on the intracellular trafficking function of neonatal Fc receptor, which rescues internalized IgGs from lysosomal degradation and recycles them back to circulation. Results: Here, we developed a novel cell-based assay to quantify recycling of monoclonal antibodies in a transwell culture system that uses a cell line that stably expresses human neonatal Fc receptor. We tested seven therapeutic antibodies and showed that the recycling output of the assay strongly correlated with the clearance in humans. Conclusion: This recycling assay has potential application as a pharmacokinetic prescreening tool to facilitate development and selection of IgG-based candidate therapeutic monoclonal antibodies.

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Real-time flow cytometry for the kinetic analysis of oncosis

Cytometry Part A ◽

10.1002/cyto.a.21022 ◽

2011 ◽

Vol 79A (3) ◽

pp. 181-191 ◽

Cited By ~ 6

Author(s):

G. Warnes ◽

S. Martins

Keyword(s):

Flow Cytometry ◽

Kinetic Analysis ◽

Real Time ◽

Time Flow

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Internalization of an Anti-Glycoprotein IIb/IIIa Antibody by HEL Cells

Thrombosis and Haemostasis ◽

10.1055/s-0038-1653767 ◽

1995 ◽

Vol 73 (02) ◽

pp. 291-296 ◽

Cited By ~ 1

Author(s):

Kenjiro Hamamoto ◽

Shosaku Nomura ◽

Masahiko Suzuki ◽

Shigetoshi Ohga ◽

Shirou Fukuhara

Keyword(s):

Flow Cytometry ◽

Monoclonal Antibodies ◽

Immunoelectron Microscopy ◽

Surface Membrane ◽

Intracellular Pool ◽

Adhesion Proteins ◽

Hel Cells

SummaryPlatelets are known to internalize monoclonal antibodies directed against the glycoprotein (GP) IIb/IIIa complex. We investigated whether an antibody directed against this complex (NNKY 2-11) was transported from the surface membrane to the intracellular pool in HEL cells. Flow cytometry showed that the percent binding of NNKY 2-11 to the surface membrane of HEL cells was decreased after incubation for 24 h compared with 1 h, while the binding of an anti-GPIb antibody (NNKY 5-5) did not change. It did not seem likely that the GP Ilb/IIIa complex antibody was shed from the surface membrane of the HEL cells during incubation, because the medium conditioned by incubation with these cells for 24 h showed almost no binding to washed platelets. In addition, immunoelectron microscopy demonstrated that GP IIb/IIIa complex antibodies were incorporated into the intracellular pool of HEL cells and were associated with alpha granules. These findings indicated that an anti-GP IIb/IIIa antibody could be internalized by megakaryocytes, as has been previously shown with platelets, suggesting that megakaryocyte GP IIb/IIIa may act as a carrier for various adhesion proteins.

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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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