Biopharmaceuticals Derived from Transgenic Plants and Animals

Báez, Julio

doi:10.1002/9783527620982.ch35

Cited by 11 publications

(8 citation statements)

References 264 publications

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“…Basic genetic engineering tools have been developed for protein production in corn and other plants, but questions of containment have delayed commercialization. 3,4 The risk of cross-pollination and resulting outcrossing of the transgenic trait to wild and domesticated relatives can be mitigated by implementing a growing strategy that provides significant distance between donor and recipient plants while carefully coordinating pollen maturation timing to avoid cross pollination with other corn plants. 4 Once harvested the grain can be stored for as much as several years before processing.…”

Section: Introductionmentioning

confidence: 99%

Purification and characterization of a transgenic corn grain‐derived recombinant collagen type I alpha 1

Zhang

Báez

Pappu³

et al. 2009

Biotechnology Progress

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Corn offers advantages as a transgenic host for producing recombinant proteins required at large volumes (1,000's of tons per year) and low cost (less than US$50/kg) by generating them as co-products of biorefining. We describe the purification and characterization of a corn grain-derived mammalian structural protein having such market characteristics: a full length recombinant collagen type I alpha 1 (rCI alpha 1) chain. Material properties of interest are gelation behavior, which would depend on as yet unverified ability of corn to carry out post-translational prolyl hydroxylation and formation of triple helical conformation. The starting material was grain where the expression of rCI alpha 1 had been directed by an embryo-specific promoter. Purification consisted of extraction at low pH followed by membrane and chromatographic steps to isolate rCI alpha 1 for characterization. The amino acid composition and immunoreactivity of CI alpha 1 was similar to that of an analogous native human CI alpha 1 and to rCI alpha 1 produced by the yeast Pichia pastoris. Tandem mass spectrometry confirmed the primary sequence of the corn-derived rCI alpha 1 with 46% coverage. Fragments of the rCI alpha 1 chains were also observed, possibly caused by endogenous plant proteases. The corn-derived rCI alpha 1 had a low level of prolyl hydroxylation (approximately 1% versus 11%) relative to animal-derived CI alpha 1 and folded into its characteristic triple-helical structure as indicated by its resistance to pepsin digestion below its melting temperature of 26(o)C. The 29 amino acid foldon fused to the C-terminus to initiate triple helix formation was not cleaved from the rCI alpha 1 chains, but could be removed by pepsin treatment.

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Section: Introductionmentioning

confidence: 99%

Purification and characterization of a transgenic corn grain‐derived recombinant collagen type I alpha 1

Zhang

Báez

Pappu³

et al. 2009

Biotechnology Progress

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“…1,2 Corn offers advantages compared with extracting proteins from animal tissues and other recombinant production systems: low-cost cultivation; easy scale-up by increased planted acreage; well-established practices for efficient harvesting, transporting, storing, and processing into multiple products; ability to fold and assemble complex proteins and some animal-like post-translational modifications; and elimination of animal-source pathogens. [3][4][5] The feasibility of commercially producing high-volume (thousands of metric tons per year) and low-cost (less than US$100/kg) recombinant proteins, depends not only on high accumulation levels in accessible parts of the plant but also on the efficient utilization of the rest of the transgenic biomass and realization of value-added coproduct revenues.…”

Section: Introductionmentioning

confidence: 99%

Fractionation of transgenic corn seed by dry and wet milling to recover recombinant collagen‐related proteins

Cheng

Glatz

Fox

et al. 2009

Biotechnology Progress

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Corn continues to be considered an attractive transgenic host for producing recombinant therapeutic and industrial proteins because of its potential for producing recombinant proteins at large volume and low cost as coproducts of corn seed-based biorefining. Efforts to reduce production costs have been primarily devoted to increasing accumulation level, optimizing protein extraction conditions, and simplifying the purification. In the present work, we evaluated two grain fractionation methods, dry milling and wet milling, to enrich two recombinant collagen-related proteins; thereby, reducing the amount and type of corn-derived impurities in subsequent protein extraction and purification steps. The two proteins were a full-length human recombinant collagen type I alpha 1(rCIalpha1) chain with telopeptides and peptide foldon to effect triple helix formation and a 44-kDa rCIalpha1 fragment. For each, approximately 60% of the rCIalpha1s in the seed was recovered in the dry-milled germ-rich fractions making up ca. 25% of the total kernel mass. For wet milling, approximately 60% of each was recovered in three fractions accounting for 20-25% of the total kernel mass. The rCIalpha1s in the dry-milled germ-rich fractions were enriched three to six times compared with the whole corn kernel, whereas the rCIalpha1s were enriched 4-10 times in selected wet-milled fractions. The recovered starch from wet milling was almost free of rCIalpha1. Therefore, it was possible to generate rCIalpha1-enriched fractions by both dry and wet milling along with rCIalpha1-free starch using wet milling. Because of its simplicity, the dry milling procedure could be accomplished on-farm thus minimizing the risk of inadvertent release of viable transgenic seeds.

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“…However, there have been advances with other hosts that they may also begin to offer valuable alternatives. There are a number of reviews available on the current state of this technology [160][161][162][163][164][165][166][167][168][169][170].…”

Section: Antibodies From Alternate Feedstocksmentioning

confidence: 99%