Stable expression of human β1,4-galactosyltransferase in plant cells modifies N-linked glycosylation patterns

Palacpac, Nirianne; Yoshida, Shohei; Sakai, Hiromi; Kimura, Yoshinobu; Fujiyama, Kazuhito; Yoshida, Takemi; Seki, Tatsuji

doi:10.1073/pnas.96.8.4692

Cited by 172 publications

(109 citation statements)

References 39 publications

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“…much more dramatically compared with WT plants than previously, when the cDNA encoding the full-length human GalT gene was introduced (12). Because MALDI-TOF MS of N-linked glycans using 2,5-dihydroxybenzoic acid can be assumed to provide an adequate reflection of the actual N-glycan makeup, it is very clear that the N-glycan pattern was more akin now to the N-glycans in a tobacco cell culture transformed with a slightly truncated cDNA lacking the first 13 amino acid residues of the cytoplasmic tail of the full-length human GalT (21,22). Both types of transgenic tobacco systems revealed a big increase in the level of high-Man and hybrid oligosaccharides and a concomitant decrease in biantennary Nglycans compared with WT plants.…”

Section: Discussionmentioning

confidence: 99%

An antibody produced in tobacco expressing a hybrid β-1,4-galactosyltransferase is essentially devoid of plant carbohydrate epitopes

Bakker

Rouwendal

Karnoup

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

126

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N-glycosylation of a mAb may have a major impact on its therapeutic merits. Here, we demonstrate that expression of a hybrid enzyme (called xylGalT), consisting of the N-terminal domain of Arabidopsis thaliana xylosyltransferase and the catalytic domain of human ␤-1,4-galactosyltransferase I (GalT), in tobacco causes a sharp reduction of N-glycans with potentially immunogenic corebound xylose (Xyl) and fucose (Fuc) residues as shown by Western blot and MALDI-TOF MS analysis. A radioallergosorbent test inhibition assay with proteins purified from leaves of WT and these transgenic tobacco plants using sera from allergic patients suggests a significant reduction of potential immunogenicity of xylGalT proteins. A mAb purified from leaves of plants expressing xylGalT displayed an N-glycan profile that featured high levels of galactose, undetectable xylose, and a trace of fucose. Hence, a transgenic plant expressing the hybrid GalT might yield more effective and safer monoclonals for therapeutic purposes than WT plants and even transgenic plants expressing the unchanged GalT.biopharmaceutical ͉ glycosylation ͉ immunogenicity

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Section: Discussionmentioning

confidence: 99%

An antibody produced in tobacco expressing a hybrid β-1,4-galactosyltransferase is essentially devoid of plant carbohydrate epitopes

Bakker

Rouwendal

Karnoup

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

126

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“…First attempts to knock in β 1-4 galactosyl transferase (GalT) were only partially successful as they rendered very heterogeneous decorations [69] [70], but the result was much better when GalT was targeted to the late Golgi. The crossing of GalT knocked in plants with ∆FT/XT mutants generated a tobacco line with a humanized glycosylation pattern.…”

Section: Deleted: Glycosilationmentioning

confidence: 99%

“…This company developed a rapid TMV-based expression system for the production of anti-idiotypic scFv antibodies against nonHodgkin's lymphoma [75]. LSB even successfully began clinical trials, but failed to Deleted: [69] Deleted: [70] Deleted: much…”

Section: Little and Fast Or Lots And Slow: Transient Versus Stable Trmentioning

confidence: 99%

Manufacturing antibodies in the plant cell

Orzáez

Granell

Blázquez

2009

Biotechnology Journal

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International audiencePlants have long been considered advantageous platforms for large-scale production of antibodies due to their low cost, scalability, and the low chances of pathogen contamination. Much effort has therefore been devoted to efficiently produce mAbs (from nanobodies to secretory antibodies) in plant cells. Several technical difficulties have been encountered and are being coped with. Improvements in production levels have been achieved by manipulation of gene expression and, more efficiently, of cell targeting and protein folding and assembly. Differences in mAb glycosylation patterns between animal and plant cells are being successfully addressed by the elimination and introduction of the appropriate enzyme activities in plant cells. Another relevant battlefield is the dichotomy between production capacity and speed. Classically, stably transformed plant lines have been proposed for large scale mAb production, whereas the use of transient expression systems has always provided production speed at the cost of scalability. However, recent advances in transient expression techniques have brought impressive yield improvements, turning speed and scalability highly compatible assets. In the era of personalized medicines, the combination of yield and speed, and the advances in glyco-engineering have made the plant cell a serious contender in the field of recombinant antibody production

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“…Nicotiana tabacum was the first plant wherein glycans were extended by typical mammalian residues through the introduction of human b1,4-galactosyltranferases [37,38]. N-glycans of antibodies produced by these plants were to a significant amount extended by terminal b1,4-galactose residues, but these glycans still carried xylose and fucose (Figure 1, Structure 6) [38].…”

Section: Glycoengineeringmentioning

confidence: 99%