Simultaneous Protein Quantitation and Glycosylation Profiling of Antigen-Specific Immunoglobulin G1 in Large Clinical Studies DOI Creative Commons
Steinar Gijze,

Anna M Wasynczuk,

Leanne P. M. van Leeuwen

et al.

Journal of Proteome Research, Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 13, 2024

Antibodies have a key role in the immune system, making their characterization essential to biomedical, biopharmaceutical, and clinical research questions. Antibody effector functions are mainly controlled by quantity, subclass, Fc glycosylation. We describe an integrated method measure these three critical dimensions simultaneously. The subclass-specific immunoglobulin G (IgG) glycosylation analysis combines immunosorbance with glycopeptide-centered LC-MS detection. For IgG1-specific quantitation, commercial, stable isotope labeled IgG1 protein standard was spiked into immunosorbent eluates. Robust quantitation achieved, relying on combination of proteotypic peptide most abundant glycopeptides, generated through proteolytic cleavage from mixture natural recombinant standard. Method performance demonstrated large coronavirus vaccination cohort at throughput 100 samples/day. LC-MS-derived, anti-SARS-CoV-2 spike concentrations ranged 10000 ng/mL correlated well clinically relevant immunoassay. Technical variation 200 times lower than biological variation; intermediate precision 44%. In conclusion, we present capable robustly simultaneously assessing antigen-specific IgG studies. This will facilitate broader understanding responses, especially important interplay among dimensions.

Language: Английский

With great power, comes great responsibility: the importance of broadly measuring Fc-mediated effector function early in the antibody development process DOI Creative Commons
Silvia Crescioli, Shashidhar S. Jatiani, Leonard Moise

et al.

mAbs, Journal Year: 2025, Volume and Issue: 17(1)

Published: Jan. 16, 2025

The field of antibody therapeutics is rapidly growing, with over 210 antibodies currently approved or in regulatory review and ~ 1,250 clinical development. Antibodies are highly versatile molecules that, strategic design their antigen-binding domain (Fab) the responsible for mediating effector functions (Fc), can be used a wide range therapeutic indications. Building on many years progress, biopharmaceutical industry now advancing innovative research development by exploring new targets formats using engineering to fine-tune tailored specific disease requirements. In addition considering target context, however, unique features each trigger diverse set Fc-mediated functions. To avoid unexpected results safety efficacy outcomes during later stages process, it crucial measure impact function early process. Given breadth deploy close interplay between Fab Fc functional domains, important conduct comprehensive evaluation an array antigen-specific biophysical cell-mediated assays. Here, we receptor properties that influence discuss implications safe efficacious therapeutics.

Language: Английский

Citations

2
Relevance of proteomics and metabolomics approaches to overview the tumorigenesis and better management of cancer DOI
Pooja Singh, Yashika Walia, Shagun Gupta

et al.

3 Biotech, Journal Year: 2025, Volume and Issue: 15(3)

Published: Feb. 11, 2025

Language: Английский

Citations

1
The importance of IgG glycosylation—What did we learn after analyzing over 100,000 individuals DOI Creative Commons
Jasminka Krištić, Gordan Lauc

Immunological Reviews, Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 4, 2024

Summary All four subclasses of immunoglobulin G (IgG) antibodies have glycan structures attached to the protein part IgG molecules. Glycans linked Fc portion are found in all antibodies, while about one‐fifth plasma also glycans Fab IgG. The IgG3 subclass is characterized by more complex glycosylation compared other subclasses. In this review, we discuss significant influence that exert on structural and functional properties We provide a comprehensive overview how composition these can affect IgG's effector functions modulating its interactions with Fcγ receptors molecules such as C1q component complement, which turn various immune responses triggered IgG, including antibody‐dependent cell‐mediated cytotoxicity (ADCC) complement‐dependent (CDC). addition, importance for efficacy therapeutics like monoclonal intravenous (IVIg) therapy discussed. Moreover, offer insights into characteristics roles derived from general population, disease‐specific, interventional studies. These studies indicate important biomarkers effectors health disease.

Language: Английский

Citations

6
Systematic analysis of Fc mutations designed to reduce binding to Fc-gamma receptors DOI Creative Commons
G Hale, Jelle De Vos,

Alastair Douglas Davy

et al.

mAbs, Journal Year: 2024, Volume and Issue: 16(1)

Published: Sept. 15, 2024

Elimination of the binding immunoglobulin Fc to gamma receptors is highly desirable for avoidance unwanted inflammatory responses therapeutic antibodies and fusion proteins. Many different approaches have been used in clinic, but they not systematically compared. We now produced a matched set anti-CD20 with subclasses variants compared their activity C1q, Fc-gamma cell-based assays. Most still significant levels one or more these assays many them impaired temperature stability corresponding wild-type antibody.

Language: Английский

Citations

5
Systematic analysis of Fc mutations designed to enhance binding to Fc-gamma receptors DOI Creative Commons
G Hale,

Alastair Douglas Davy,

Ian B. Wilkinson

et al.

mAbs, Journal Year: 2024, Volume and Issue: 16(1)

Published: Sept. 22, 2024

A critical attribute of therapeutic antibodies is their ability to engage with humoral or cellular effector mechanisms, and this depends on the Fc region bind complement (C1q) receptors. Investigators have sought optimize these effects by engineering a greater lesser extent individual Different approaches been used in clinic, but they not systematically compared. We now produced matched set anti-CD20 representing range variants compared activity cell-based assays for complement-dependent cytotoxicity, antibody-dependent cell-mediated phagocytosis using also thermal stability differential scanning fluorimetry (DSF). The results reveal spectrum activities which may be appropriate different applications.

Language: Английский

Citations

5
Complement‐targeted therapeutics: Are we there yet, or just getting started? DOI Creative Commons
Daniel Ricklin

European Journal of Immunology, Journal Year: 2024, Volume and Issue: 54(12)

Published: Sept. 12, 2024

Therapeutic interventions in the complement system, a key immune-inflammatory mediator and contributor to broad range of clinical conditions, have long been considered important yet challenging or even unfeasible achieve. Almost 20 years ago, spark was lit demonstrating commercial viability complement-targeted therapies. Since then, field has experienced an impressive expansion targeted indications available treatment modalities. Currently, dozen distinct complement-specific therapeutics covering several intervention points are clinic, benefiting patients suffering from eight disorders, not counting numerous trials off-label uses. Observing this rapid rise therapy obscurity mainstream with amazement, one might ask whether peak development now reached will continue marching on new heights. This review looks at milestones drug discovery achieved so far, surveys currently approved entities indications, ventures glimpse into future advancements come.

Language: Английский

Citations

4
Fc Gamma Receptor Polymorphisms in Antibody Therapy: Implications for Bioassay Development to Enhance Product Quality DOI Creative Commons
Julianne D. Twomey, S. Elizabeth George,

Bao-Lin Zhang

et al.

Antibody Therapeutics, Journal Year: 2025, Volume and Issue: 8(2), P. 87 - 98

Published: Jan. 21, 2025

Abstract The effectiveness of therapeutic antibodies is often associated with their Fc-mediated effector functions, such as antibody-dependent cellular cytotoxicity and phagocytosis. These functions rely on interactions between Fc gamma receptors (FcγRs) immune cells the region antibodies. Genetic variations in these receptors, known FcγR polymorphisms, can influence outcomes by altering receptor expression levels, affinity, function. This review examines impact polymorphisms antibody therapy, emphasizing role developing optimizing functional bioassays to assess product quality. Understanding essential for refining bioassays, which are crucial accurately characterizing products ensuring consistency manufacturing processes.

Language: Английский

Citations

0
Complement-coagulation crosstalk in idiopathic membranous nephropathy: The potential pathogenesis and therapeutic perspective DOI Creative Commons
Zhirong Liu, Wei Liang, Yangbin Pan

et al.

Autoimmunity Reviews, Journal Year: 2025, Volume and Issue: unknown, P. 103763 - 103763

Published: Feb. 1, 2025

Idiopathic membranous nephropathy (IMN) is a glomerular disease that prevalent in elderly males. The pathogenesis of IMN includes abnormal autoimmunity and complement activation, both which leading to the damage filtration structure. Meanwhile, due pathological changes kidney, certain coagulation-related proteins are leaked from urine, resulting imbalance coagulation homeostasis. Recent studies have indicated interaction between systems, while aberration common IMN. In this review, we summarize subsistent underlying ensue complement-coagulation crosstalk present emerging evidence evolving field.

Language: Английский

Citations

0
N-glycosylation signature and its relevance in cardiovascular immunometabolism DOI Creative Commons
Monika Svecla, Ruifang Li‐Gao, David Falck

et al.

Vascular Pharmacology, Journal Year: 2025, Volume and Issue: unknown, P. 107474 - 107474

Published: Feb. 1, 2025

Language: Английский

Citations

0
Enhancing activity of FcαRI-bispecific antibodies using glycoengineering DOI Creative Commons
Celine Angeli Natascha Sewnath, Timon Damelang, Arthur E. H. Bentlage

et al.

The Journal of Immunology, Journal Year: 2025, Volume and Issue: unknown

Published: March 28, 2025

Macrophages and natural killer (NK) cells can effectively kill tumor in the presence of anti-cancer IgG monoclonal antibodies (mAbs), but neutrophils are less effective. We previously showed that IgG1 bispecific (BsAb), which target IgA Fc receptor (FcαRI, CD89) a associated antigen induce effective neutrophil recruitment cell killing vivo. Here we investigated if efficacy an anti-EGFR (CetuximAb)/FcαRI-bispecific antibody could be further improved by implementing glycoengineering IgG-Fc, aimed at increasing FcγRIIIa/b binding and/or complement activity. afucosylation was introduced to enhance antibody-dependent cellular cytotoxicity (ADCC) FcγRIIIa on NK/macrophages, also reduce neutrophil-mediated ADCC through their GPI-linked FcγRIIIb. galactylation found hexamerization thereby dependent (CDC). Low fucosylated BsAbs moderately increased NK cell-mediated killing, did not affect nor phagocytosis macrophages. Glycoengineering these EGFR-specific BsAb, normally devoid CDC-activity, enable activities. In conclusion, glycoengineered FcαRI had little effect or macrophage mediated killing.

Language: Английский

Citations

0