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Human protein C (hPC) is glycosylated at three Asn-X-Ser/Thr and one atypical Asn-X-Cys sequons. We have characterized the micro- and macro-heterogeneity of plasma-derived hPC and compared the glycosylation features with recombinant protein C (tg-PC) produced in a transgenic pig bioreactor from two animals having approximately tenfold different expression(More)
Glycosylation of recombinant proteins is of particular importance because it can play significant roles in the clinical properties of the glycoprotein. In this work, the N-glycan structures of recombinant human Factor IX (tg-FIX) produced in the transgenic pig mammary gland were determined. The majority of the N-glycans of transgenic pig-derived Factor IX(More)
Appropriate glycosylation of recombinant therapeutic glycoproteins has been emphasized in biopharmaceutical industries because the carbohydrate component can affect safety, efficacy, and consistency of the glycoproteins. Reliable quantification methods are essential to ensure consistency of their products with respect to glycosylation, particularly(More)
Both the low animal cell density of bioreactors and their ability to post-translationally process recombinant factor IX (rFIX) limit hemophilia B therapy to <20% of the world's population. We used transgenic pigs to make rFIX in milk at about 3,000-fold higher output than provided by industrial bioreactors. However, this resulted in incomplete(More)
Exploring the phase during wound healing when fibrinogen (FBG) and fibronectin (FN) interact forming a small thin layer for cells to migrate and proliferate at the injury site is necessary for the primary building blocks and initial stages of skin recovery. The aim of this thesis is to illustrate that administrating a reasonable amount of (FN) and FBG, in a(More)
A fibrinogen-fibronectin complex (γγ'pdFI-pdFN) was purified from normal human plasma using a sequence of cryoprecipitation, ammonium sulfate fractionation, and DEAE Sepharose chromatography. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) under reducing condition showed both a 1:1 stoichiometric ratio of fibrinogen (FI) to fibronectin(More)
Pichia pastoris has demonstrated the ability to express high levels of recombinant heterologous proteins. Protein expression is enhanced during fermentation at high cell density. However, the level of expression is mainly regulated by fermentation operation factors. This research is directed to investigate the effect of methanol growth rate, temperature and(More)
This study uses high-pressure size exclusion chromatography (HPSEC) to quantify divalent metal ion (X 2+)-induced compaction found in vitamin K dependent (VKD) proteins. Multiple X 2+ binding sites formed by the presence of up to 12-carboxyglutamic acid residues (Gla) are present in plasma-derived (pd-) and recombinant (r-) Factor IX (FIX). Analytical(More)
Noncompressible truncal hemorrhage and brain injury currently account for most early mortality of warfighters on the battlefield. There is no effective treatment for noncompressible truncal hemorrhage, other than rapid evacuation to a surgical facility. The availability of an effective field treatment for noncompressible truncal hemorrhage could increase(More)