N- and O-glycans of recombinant human C1 inhibitor expressed in the milk of transgenic rabbits

Koles, Kate; Berkel, Patrick H.C. van; Pieper, Frank; Nuijens, Jan H.; Mannesse, Maurice; Vliegenthart, Johannes F.G.; Kamerling, Johannis P.

doi:10.1093/glycob/cwh010

Cited by 91 publications

(72 citation statements)

References 48 publications

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“…To date several proteins have been expressed in the milk of transgenic animals in quantities ranging from a few micrograms to several grams per liter. Some examples are: human lactoferrin in mice [2] and cows [3]; a1-antitrypsin [4] and C1 inhibitor [5] in rabbits; human anti-thrombin [6] and human tissue-type plasminogen activator [7,8] in goats; human Factor VIII [9] and human Protein C [10] in pigs; human a-glucosidase in mice [11] and rabbits [12] and human erythropoietin in rabbits [13] and pigs [14]. However, the functionality and specificity of mammary gland glycosylation is, as yet, incompletely understood in comparison with other mammalian cell lines.…”

mentioning

confidence: 99%

Monosialylated biantennary N-glycoforms containing GalNAc–GlcNAc antennae predominate when human EPO is expressed in goat milk

Montesino

Toledo

Sánchez

et al. 2008

Archives of Biochemistry and Biophysics

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Recently, our group reported the expression of recombinant human erythropoietin in goat milk (rhEPO-milk) as well as in the mammary epithelial cell line GMGE (EPO-GMGE) by cell culture using the adenoviral transduction system. N-Glycosylation characterization of rhEPO-milk by Normal-Phase HPLC profiling of the fluorophore, 4-aminobenzoic acid-labeled enzymatically released N-glycan pool from rhEPO-goat milk, combined with MALDI, ESI-MS and LC/MS, revealed that low branched, core-fucosylated, N-glycans predominate. The labeled N-glycans were separated into neutral and charged fractions by anion exchange chromatography and the charged N-glycans were found to be mostly a2,6-monosialylated with Neu5Ac or Neu5Gc in a ratio of 1:1. Unlike the N-glycans from rhEPO produced in CHO cells, where the glycans are multiantennary highly sialylated, core-fucosylated oligosaccahrides, or even in the goat mammary gland epithelial cell line cultured in vitro in which multiantennary, core-and outer-arm fucosylated, monosialylated Nglycans are the most abundant species, a large proportion of the N-glycans from rhEPO-milk were monosialylated, biantennary, antennae mostly terminating with the more unusual GalNAc-GlcNAc motive and without outer-arm fucosylation. These findings, emphasizing the difference in the N-glycan repertoire between the rhEPO-milk and EPO-GMGE, are consistent with the principle that glycosylation is cell-type dependent and that the cell environment is crucial as well.

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mentioning

confidence: 99%

Monosialylated biantennary N-glycoforms containing GalNAc–GlcNAc antennae predominate when human EPO is expressed in goat milk

Montesino

Toledo

Sánchez

et al. 2008

Archives of Biochemistry and Biophysics

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“…The proteins lacking sialic acid generally have an excess of terminal mannose which favours their rapid elimination from the blood. Interestingly, the protein C inhibitor [33] and a coagulation blood factor (unpublished data) secreted in rabbit milk did not contain fucose. This suggests that fucosylation is poorly active in the rabbit mammary gland.…”

Section: Production Of Pharmaceutical Proteins In Milkmentioning

confidence: 98%

“…Rabbit and pig milking can be achieved with no particular difficulty using adapted machines. Rabbits and pigs appeared more able than ruminants to perform some posttranslational modifications as judged by the examination of a few proteins such as human protein C [36] and human protein C inhibitor [33]. This is particularly the case for glycosylation.…”

Section: Production Of Pharmaceutical Proteins In Milkmentioning

confidence: 99%

“…This is particularly the case for glycosylation. Rabbits add almost quantitatively sialic acid under the NANA form (N-acetylneuranumic acid) to protein C inhibitor [33], whereas goats are unable to sialylate completely antithrombin III and about half of the sialic acid is under the NGNA form (N-glycosylneuranumic acid) [37]. The latter form is not present in human proteins and its presence in recombinant proteins could generate the formation of antibodies by patients.…”

Section: Production Of Pharmaceutical Proteins In Milkmentioning

confidence: 99%

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Preparation of recombinant proteins in milk to improve human and animal health

Soler¹,

Thépot²,

Rival‐Gervier³

et al. 2006

Reprod. Nutr. Dev.

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-Milk is a very abundant source of proteins for animal and human consumption. Milk composition can be modified using transgenesis, including exogenous gene addition and endogenous gene inactivation. The study of milk protein genes has provided researchers with regulatory regions capable of efficiently and specifically driving the expression of foreign genes in milk. The projects underway are aimed at modifying milk composition, improving its nutritional value, reducing mammary infections, providing consumers with antipathogen proteins and preparing purified recombinant proteins for pharmaceutical use. The present paper summarises the current progress in this field. milk / recombinant proteins / animal and human health

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“…A new approach for the development of a human C1 inhibitor for treatment of HAE attacks was developed by a second Dutch company, Pharming[20]. This company pioneered the concept of isolating the human gene for a plasma protein and fusing it to a bovine casein promoter.…”

Section: Recombinant C1 Inhibitormentioning

confidence: 99%

Recombinant and Plasma-Purified Human C1 Inhibitor for the Treatment of Hereditary Angioedema

Frank¹

2010

World Allergy Organization Journal

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BackgroundAgents for prophylaxis of hereditary angioedema (HAE) have been available in the United States for several decades, but their usefulness is limited by side effects and they cannot be used at all in some patients. No agents have been available in the United States to specifically treat acute attacks. HAE types I and II are associated with low functional levels of C1 inhibitor, and evidence accumulated over decades suggests that intravenous infusion of C1 inhibitor is useful for terminating angioedema attacks and for prophylaxis.Key PointsC1 inhibitor derived from pooled human plasma has been available for decades in Europe, and 2 preparations have been recently introduced into the United States. Both have been efficacious in carefully controlled double-blind studies. One preparation, Cinryze, was approved by the U.S. Food & Drug Administration (FDA) for prophylaxis of HAE attacks in October 2008, and the second, Berinert, was approved by the FDA for treatment of acute attacks in October 2009. A third preparation, the recombinant human C1 inhibitor Rhucin, is completing clinical trials. Although not yet approved by the FDA, preliminary results made available suggest that Rhucin, too, is effective in terminating attacks of HAE.ConclusionsAmericans will now have access to effective acute and prophylactic treatments with C1 inhibitor for hereditary angioedema.

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N- and O-glycans of recombinant human C1 inhibitor expressed in the milk of transgenic rabbits

Cited by 91 publications

References 48 publications

Monosialylated biantennary N-glycoforms containing GalNAc–GlcNAc antennae predominate when human EPO is expressed in goat milk

Monosialylated biantennary N-glycoforms containing GalNAc–GlcNAc antennae predominate when human EPO is expressed in goat milk

Preparation of recombinant proteins in milk to improve human and animal health

Recombinant and Plasma-Purified Human C1 Inhibitor for the Treatment of Hereditary Angioedema

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