IgG1 versus IgG3: influence of antibody-specificity and allotypic variance on virus neutralization efficacy DOI Creative Commons
Somanath Kallolimath, Lin Sun, Roman Palt

и другие.

Frontiers in Immunology, Год журнала: 2024, Номер 15

Опубликована: Окт. 24, 2024

Despite the unique advantages of IgG3 over other IgG subclasses, such as mediating enhanced effector functions and increased flexibility in antigen binding due to a long hinge region, therapeutic potential remains largely unexplored. This may be attributed difficulties recombinant expression reduced plasma half-life most allotypes. Here, we report plant two SARS-CoV-2 neutralizing monoclonal antibodies (mAbs) that exhibit high (P5C3) low (H4) binding. P5C3 H4-IgG1 mAbs were subclass-switched formats, designed for efficient production PK values, carrying three allotypic variations, referred -WT, -H, -KVH. A total eight produced glycoengineered plants synthesize fucose-free complex N-glycans with great homogeneity. Antigen, IgG-FcγR immune complement studies demonstrated similar activities all mAbs. In accordance, Abs showed minor alterations neutralization (NT) antibody-dependent cell-mediated virus inhibition (ADCVI). Clear functional differences observed between H4 variants superior ADCVI NT potencies H. Our comparative study demonstrates an variant Fc domain equivalent or compared IgG3-WT, but stability values ​​of IgG1. data also demonstrate both variability antibody specificity are important fine-tuning activities, information development future therapeutics.

Язык: Английский

Differential prolyl hydroxylation by six Physcomitrella prolyl-4 hydroxylases DOI Creative Commons
Christine Rempfer, Sebastian N. W. Hoernstein, Nico van Gessel

и другие.

Computational and Structural Biotechnology Journal, Год журнала: 2024, Номер 23, С. 2580 - 2594

Опубликована: Июнь 14, 2024

Hydroxylation of prolines to 4-trans-hydroxyproline (Hyp) is mediated by prolyl-4 hydroxylases (P4Hs). In plants, Hyps occur in Hydroxyproline-rich glycoproteins (HRGPs), and are frequently O-glycosylated. While both modifications important, e.g. for cell wall stability, they undesired plant-made pharmaceuticals. Sequence motifs prolyl-hydroxylation were proposed but did not include data from mosses, such as Physcomitrella. We identified six moss P4Hs phylogenetic reconstruction. Our analysis 73 24 secretory proteins multiple mass spectrometry datasets revealed that near other prolines, alanine, serine, threonine valine preferentially hydroxylated. About 95 % predictable with combined established methods. our data, AOV was the most frequent pattern. A combination 443 AlphaFold models MS 3000 found mainly on protein surfaces disordered regions. Moss-produced human erythropoietin (EPO) exhibited O-glycosylation arabinose chains two Hyps. This modification significantly reduced a p4h1 knock-out (KO) Physcomitrella mutant. Quantitative proteomics different p4h mutants specific changes amounts, modified pattern, suggesting differential function P4Hs. RT-PCR effect single KOs expression five genes, partial compensation mutation. AlphaFold-Multimer P4H1 its target EPO peptide superposed crystal structure Chlamydomonas suggested significant amino acids active centre enzyme differences between

Язык: Английский

Процитировано

1
IgG1 versus IgG3: influence of antibody-specificity and allotypic variance on virus neutralization efficacy DOI Creative Commons
Somanath Kallolimath, Lin Sun, Roman Palt

и другие.

Frontiers in Immunology, Год журнала: 2024, Номер 15

Опубликована: Окт. 24, 2024

Despite the unique advantages of IgG3 over other IgG subclasses, such as mediating enhanced effector functions and increased flexibility in antigen binding due to a long hinge region, therapeutic potential remains largely unexplored. This may be attributed difficulties recombinant expression reduced plasma half-life most allotypes. Here, we report plant two SARS-CoV-2 neutralizing monoclonal antibodies (mAbs) that exhibit high (P5C3) low (H4) binding. P5C3 H4-IgG1 mAbs were subclass-switched formats, designed for efficient production PK values, carrying three allotypic variations, referred -WT, -H, -KVH. A total eight produced glycoengineered plants synthesize fucose-free complex N-glycans with great homogeneity. Antigen, IgG-FcγR immune complement studies demonstrated similar activities all mAbs. In accordance, Abs showed minor alterations neutralization (NT) antibody-dependent cell-mediated virus inhibition (ADCVI). Clear functional differences observed between H4 variants superior ADCVI NT potencies H. Our comparative study demonstrates an variant Fc domain equivalent or compared IgG3-WT, but stability values ​​of IgG1. data also demonstrate both variability antibody specificity are important fine-tuning activities, information development future therapeutics.

Язык: Английский

Процитировано

0