Glycoengineered antibodies: towards the next-generation of immunotherapeutics

Mastrangeli, Renato; Palinsky, Wolf; Bierau, Horst

doi:10.1093/glycob/cwy092

Cited by 68 publications

(47 citation statements)

References 102 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The results showed that field-grown tobacco expressing a recombinant protein were capable of achieving an order of magnitude reduction in costs compared to traditional cell culture methods [44]. In addition, as the glycosylation state of antibody therapeutics is crucial for their function [45,46], plants are uniquely suited to produce RIC vaccines. Antibodies made in glycoengineered plants have demonstrated improved C1q, FccRI, and FccRIIIa binding, leading to improved antibody-dependent cellular cytotoxicity, complement-dependent cytotoxicity, and overall increased potency compared with commercial antibodies made in mammalian cells [47,48] Here, we find that fully assembled and highly expressed (Figs.…”

Section: Discussionmentioning

confidence: 99%

Codelivery of improved immune complex and virus-like particle vaccines containing Zika virus envelope domain III synergistically enhances immunogenicity

Diamos

Pardhe

Sun

et al. 2020

Vaccine

View full text Add to dashboard Cite

a b s t r a c tZika virus (ZIKV) reemergence poses a significant health threat especially due to its risks to fetal development, necessitating safe and effective vaccines that can protect pregnant women. Zika envelope domain III (ZE3) has been identified as a safe and effective vaccine candidate, however it is poorly immunogenic. We previously showed that plant-made recombinant immune complex (RIC) vaccines are a robust platform to improve the immunogenicity of weak antigens. In this study, we altered the antigen fusion site on the RIC platform to accommodate N-terminal fusion to the IgG heavy chain (N-RIC), and thus a wider range of antigens, with a resulting 40% improvement in RIC expression over the normal C-terminal fusion (C-RIC). Both types of RICs containing ZE3 were efficiently assembled in plants and purified to >95% homogeneity with a simple one-step purification. Both ZE3 RICs strongly bound complement receptor C1q and elicited strong ZE3-specific antibody titers that correlated with ZIKV neutralization. When either N-RIC or C-RIC was codelivered with plant-produced hepatitis B core (HBc) virus-like particles (VLP) displaying ZE3, the combination elicited 5-fold greater antibody titers (>1,000,000) and more strongly neutralized ZIKV than either RICs or VLPs alone, after only two doses without adjuvant. These findings demonstrate that antigens that require a free N-terminus for optimal antigen display can now be used with the RIC system, and that plant-made RICs and VLPs are highly effective vaccines targeting ZE3. Thus, the RIC platform can be more generally applied to a wider variety of antigens.

show abstract

Section: Discussionmentioning

confidence: 99%

Codelivery of improved immune complex and virus-like particle vaccines containing Zika virus envelope domain III synergistically enhances immunogenicity

Diamos

Pardhe

Sun

et al. 2020

Vaccine

View full text Add to dashboard Cite

show abstract

“…As indicated by our results, the final product might be redesigned in a precise manner, reducing the need of host cell engineering and variations arising from the manufacturing process. Moreover, this technique offers speedy production with free choice of the expression system, 15 although its introduction would certainly require additional processing steps into existing operations, such as monitoring of product purity to exclude the risk of additional contaminants, contributing itself to the overall manufacturing costs and a new regulatory framework.…”

Section: Discussionmentioning

confidence: 99%

“…14 This approach increases efficacy and safety of the final product from the health-care perspective, and facilities process development offering host expression flexibility and lower batch-to-batch inconsistency. 15 To better understand the impact of N-glycosylation on mAb-FcγR binding affinity, a variety of biophysical analytical tools such as surface plasmon resonance (SPR), 16 isothermal titration calorimetry (ITC), 17 nuclear magnetic resonance (NMR) 18 and X-ray crystallography are used. 19 Among these, SPR is often considered the "gold standard" in protein-protein interaction studies.…”

Section: Introductionmentioning

confidence: 99%

Interaction analysis of glycoengineered antibodies with CD16a: a native mass spectrometry approach

Hajduk

Brunner

Malik

et al. 2020

mAbs

View full text Add to dashboard Cite

2020) Interaction analysis of glycoengineered antibodies with CD16a: a native mass spectrometry approach, mAbs, 12:1, 1736975, ABSTRACTMinor changes in the quality of biologically manufactured monoclonal antibodies (mAbs) can affect their bioactivity and efficacy. One of the most important variations concerns the N-glycosylation pattern, which directly affects an anti-tumor mechanism called antibody-dependent cell-meditated cytotoxicity (ADCC). Thus, careful engineering of mAbs is expected to enhance both protein-receptor binding and ADCC. The specific aim of this study is to evaluate the influence of terminal carbohydrates within the Fc region on the interaction with the FcγRIIIa/CD16a receptor in native and label-free conditions. The single mAb molecule comprises variants with minimal and maximal galactosylation, as well as α2,3 and α2,6-sialic acid isomers. Here, we apply native electrospray ionization mass spectrometry to determine the solution-phase antibody-receptor equilibria and by using temperature-controlled nanoelectrospray, a thermal stability of the complex is examined. Based on these, we prove that the galactosylation of a fucosylated Fc region increases the binding to CD16a 1.5-fold when compared with the nongalactosylated variant. The α2,6-sialylation has no significant effect on the binding, whereas the α2,3-sialylation decreases it 1.72-fold. In line with expectation, the galactoslylated and α2,6-sialylated mAb:CD16a complex exhibit higher thermal stability when measured in the temperature gradient from 20 to 50°C. The similar binding pattern is observed based on surface plasmon resonance analysis and immunofluorescence staining using natural killer cells. The results of our study provide new insight into N-glycosylation-based interaction of the mAb:CD16a complex.

show abstract

“…For example, we could show that Fc fragments containing complex biantennary glycans attached to both the N- and C-terminal ends of the Fc could inhibit influenza A-mediated agglutination of human erythrocytes (24). The aim of the current study was to reveal possible variation in functional glycosylation related to differences in two host cell lines, CHO-K1 and HEK 293-F, particularly as antibodies and Fc fusions are the fastest growing therapeutic class in the pharmaceutical industry (26, 37, 38).…”

Section: Discussionmentioning

confidence: 99%

Choice of host cell line is essential for the functional glycosylation of the fragment crystallizable (Fc) region of human IgG1 inhibitors of influenza B viruses

Blundell

Dell

et al. 2019

Preprint

View full text Add to dashboard Cite

Antibodies are glycoproteins that carry a conserved N-linked carbohydrate attached to the Fc, whose presence and fine structure profoundly impacts on their in vivo immunogenicity, pharmacokinetics and functional attributes. The host cell line used to produce IgG has a major impact on this glycosylation, as different systems express different glycosylation enzymes and transporters that contribute to the specificity and heterogeneity of the final IgG-Fc glycosylation profile. Here we compare two panels of glycan-adapted IgG1-Fc mutants expressed in either the HEK 293-F or CHO-K1 systems. We show that the types of N-linked glycans between matched pairs of Fc mutants vary significantly, and in particular with respect to sialylation. These cell line effects on glycosylation profoundly influence the ability of the engineered Fcs to interact with either human or pathogen receptors. For example, we describe Fc mutants that potently disrupted influenza B-mediated agglutination of human erythrocytes when expressed in CHO-K1 but not in HEK 293-F cells.

show abstract

Glycoengineered antibodies: towards the next-generation of immunotherapeutics

Cited by 68 publications

References 102 publications

Codelivery of improved immune complex and virus-like particle vaccines containing Zika virus envelope domain III synergistically enhances immunogenicity

Codelivery of improved immune complex and virus-like particle vaccines containing Zika virus envelope domain III synergistically enhances immunogenicity

Interaction analysis of glycoengineered antibodies with CD16a: a native mass spectrometry approach

Choice of host cell line is essential for the functional glycosylation of the fragment crystallizable (Fc) region of human IgG1 inhibitors of influenza B viruses

Contact Info

Product

Resources

About