Purification of the Antihemophilic Factor by Gel Filtration on Agarose

1969 ◽  
Vol 22 (03) ◽  
pp. 577-583 ◽  
Author(s):  
M.M.P Paulssen ◽  
A.C.M.G.B Wouterlood ◽  
H.L.M.A Scheffers
Keyword(s):  
Factor Viii ◽  
Plasma Proteins ◽  
Large Scale ◽  
Gel Filtration ◽  
Large Scale Preparation ◽  
Scale Preparation ◽  
Antihemophilic Factor

SummaryFactor VIII can be isolated from plasma proteins, including fibrinogen by chromatography on agarose. The best results were obtained with Sepharose 6B. Large scale preparation is also possible when cryoprecipitate is separated by chromatography. In most fractions containing factor VIII a turbidity is observed which may be due to the presence of chylomicrons.The purified factor VIII was active in vivo as well as in vitro.

Large-Scale Preparation of Recombinant Ovine Prolactin and Determination of Its in Vitro and in Vivo Activity

2001 ◽  
Vol 22 (3) ◽  
pp. 489-496 ◽  
Author(s):  
Haim Leibovich ◽  
Nina Raver ◽  
Asael Herman ◽  
Ewa L. Gregoraszczuk ◽  
Elisha Gootwine ◽  
...  
Keyword(s):  
Large Scale ◽  
Large Scale Preparation ◽  
Scale Preparation ◽  
Ovine Prolactin

Large-Scale Preparation of a Highly Purified Solvent-Detergent Treated Factor VIII Concentrate

Vox Sanguinis ◽  
1991 ◽  
Vol 60 (3) ◽  
pp. 141-147
Author(s):  
Milan Wickerhauser ◽  
Leigh Charamella ◽  
Robert Myers ◽  
Louise Simon ◽  
William Nummy ◽  
...  
Keyword(s):  
Factor Viii ◽  
Large Scale ◽  
Large Scale Preparation ◽  
Scale Preparation

Inhibition of Human Platelet Aggregation by the Proteolytic Effect of Streptokinase (SK). Role of the Factor VIII Related Antigen

1975 ◽  
Author(s):  
R. Castillo ◽  
S. Maragall ◽  
J. A. Guisasola ◽  
F. Casals ◽  
C. Ruiz ◽  
...  
Keyword(s):  
Platelet Aggregation ◽  
Factor Viii ◽  
Platelet Rich Plasma ◽  
Gel Filtration ◽  
Inhibitory Effect ◽  
Factor Viii Related Antigen ◽  
Human Factor Viii ◽  
Induced Aggregation

Defective ADP-induced platelet aggregation has been observed in patients treated with streptokinase. This same effect appears “in vitro” when adding SK to platelet rich plasma (PRP). Classic hemophilia and normal platelet poor plasmas (PPP) treated with SK inhibit the aggregation of washed platelets; plasmin-treated normal human serum also shows an inhibitory effect on platelet aggregation. However, von Willebrand SK-treated plasmas do not inhibit the aggregation of washed platelets. The same results appear when plasmas are previously treated with a rabbit antibody to human factor VIII.This confirms that the antiaggregating effect is mainly linked to the digested factor VIII related antigen.The inhibition of ADP-induced platelet aggregation has been proved in gel filtration-isolated and washed platelets from SK-treated PRP.Defective ristocetin-induced platelet aggregation has also been observed- This action does not appear in washed platelets from SK-treated PRP in presence of normal PPP, but it does in presence of SK-treated PPP, which suggests that the inhibition of the ristocetin-induced aggregation is due to the lack of factor VIII and not to the factor VIII-related products.Heparin, either “in vivo” or “in vitro”, has corrected the antiaggregating effect of SK.

scholarly journals Ostrich crystallins. Structural characterization of δ-crystallin with enzymic activity

1991 ◽  
Vol 273 (2) ◽  
pp. 295-300 ◽  
Author(s):  
S H Chiou ◽  
C H Lo ◽  
C Y Chang ◽  
T Itoh ◽  
H Kaji ◽  
...  
Keyword(s):  
Large Scale ◽  
Gel Filtration ◽  
Helical Structure ◽  
Enzymic Activity ◽  
Lens Crystallins ◽  
Large Scale Preparation ◽  
Helical Content ◽  
Lyase Activity ◽  
Molecular Masses ◽  
Scale Preparation

Lens crystallins from the African ostrich (Struthio camelus) were isolated and characterized. Four crystallin fractions corresponding to alpha-, delta/beta- and beta-crystallins similar to those of duck crystallins were isolated, but epsilon-crystallin was found to be absent. The native molecular masses and subunit structures of the purified fractions were analysed by gel filtration. SDS/PAGE and isoelectric focusing, revealing various extents of heterogeneity in each orthologous crystallin class. An ion-exchange chromatographic method was used for the large-scale preparation of delta-crystallin suitable for structural and enzymic studies. It was unexpectedly found that the purified native delta-crystallin of ostrich lens possessed high argininosuccinate lyase activity, in contrast with chicken delta-crystallin. The c.d. spectra indicated a predominant beta-sheet structure in alpha- and beta-crystallins, and a significant contribution of alpha-helical structure in the delta-crystallin fraction. The estimate of secondary structures from c.d. spectroscopy for each crystallin class bears a resemblance to that of duck crystallins, except that ostrich delta-crystallin possesses much less helical content than duck delta-crystallin. Comparison of crystallin compositions and structures from aquatic and terrestrial birds revealed distinct differences.

Fe-Based Single-Atom Nanozyme with Superior Peroxidase-Mimicking Activity for Enhanced Ultrasensitive Biosensing

2021 ◽  
Vol 21 (12) ◽  
pp. 6126-6134
Author(s):  
Lili Chi ◽  
Yuetong Zhang ◽  
Yusheng Hua ◽  
Qiqi Xu ◽  
Mingzhu Lv ◽  
...  
Keyword(s):  
Active Sites ◽  
Large Scale ◽  
Low Cost ◽  
Single Atom ◽  
Specificity And Sensitivity ◽  
Large Scale Preparation ◽  
Species Formation ◽  
Biomedical Diagnosis ◽  
Scale Preparation

Nanomaterials with intrinsic enzyme-mimicking characteristics, refered to as nanozymes, have become a hot research topic owing to their unique advantages of comparative low cost, high stability and large-scale preparation. Among them, Single-atom nanozymes (SAzymes), as novel nanozymes with abundant atomically dispersed active sites, have caused specific attention in the development of nanozymes for their remarkable catalytic activities, maximum atomic utilization and excellent selectivity, the homogeneous catalytic sites and clear catalytic mechanisms. Herein, a novel single-atom nanozyme based on Fe(III)-doped polydiaminopyridine nanofusiforms (Fe-PDAP SAzyme) was successfully proposed via facile oxidation polymerization strategy. With well-defined coordination structure and abundant Fe-Nx active sites similar to natural metalloproteases, the Fe-PDAP SAzyme exhibits superior peroxidase-like activity by efficiently decomposing H2O2 for hydroxyl radical (.OH) species formation. Based on their superior peroxidase-like activity, colorimetric biosensing of H2O2 and glucose in vitro was performed by using a typical 3,3,5,5-tetramethylbenzidine through a multienzyme biocatalytic cascade platform, exhibiting the superior specificity and sensitivity. This work not only provides a novel promising SAzyme-based biosensor but also paves an avenue for evaluating enzyme activity and broadens the application of other nanozyme-based biosensors in the fields of biomedical diagnosis.

Large-scale preparation of sialyl-Tn antigen-containing peptides from mucin-like glycoproteins in boar seminal gel

2021 ◽  
Author(s):  
Ryota Takeuchi ◽  
Megumi Maeda ◽  
Miran Nakano ◽  
Hiroaki Funahashi ◽  
Yoshinobu Kimura
Keyword(s):  
Tumor Antigen ◽  
Large Scale ◽  
Gel Filtration ◽  
New Method ◽  
Tn Antigen ◽  
Large Scale Preparation ◽  
Scale Preparation ◽  
Sialyl Tn Antigen ◽  
Unused Resource ◽  
Sialyl Tn

Abstract Sialyl-Tn antigen, a tumor antigen, is a valuable ligand for the purification of proteins that specifically bind to it. Here, we developed a new method for the preparation of large amounts of sialyl-Tn antigen-containing peptides from an unused resource, boar seminal gel. The glycopeptides were prepared from the actinase E digests by a combination of gel filtration and hydrophilic partitioning.

Preparation of a partially desialylated human chorionic gonadotrophin (hCG) and its use for induction of ovulation after ovarian stimulation with human menopausal gonadotrophin

1980 ◽  
Vol 95 (2) ◽  
pp. 232-236 ◽  
Author(s):  
P. G. Crosignani ◽  
P. Donini ◽  
G. C. Lombroso ◽  
S. Donini ◽  
A. Caccamo ◽  
...  
Keyword(s):  
Large Scale ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Human Beings ◽  
Large Scale Preparation ◽  
Human Menopausal Gonadotrophin ◽  
Scale Preparation ◽  
Human Use ◽  
Sepharose 4B

Abstract. A method for the large scale preparation of partially desialylated human chorionic gonadotrophin suitable for human use is reported. To obtain the desired grade of desialylation and to avoid the presence of the enzyme in the modified hormone, neuraminidase coupled to Sepharose 4B was used. The preparation showed to be active in vitro (OAAD and SVW tests) and its half-life was found to be 13 min in the rat and 75 min in human beings. This desialo hCG proved to be effective in inducing ovulation in amenorrhoeic women. Among 39 induced cycles 31 ovulations and 5 pregnancies occurred.

scholarly journals The Large Scale Preparation of Clottable Fibrinogen-Free, High Purity, High Potency Factor VIII Concentrate

1977 ◽  
Author(s):  
L.F. Fekete ◽  
W.L. Wilson ◽  
R.L. Bick
Keyword(s):  
Factor Viii ◽  
High Speed ◽  
Large Scale ◽  
Degradation Products ◽  
Electrophoretic Analysis ◽  
High Molecular Weight Polymer ◽  
Molecular Weight Polymer ◽  
Theoretical Yield ◽  
Large Scale Preparation ◽  
Scale Preparation

Crude Factor VIII was initially extracted from plasma as cryoprecipitate. This crude concentrate was treated with high molecular weight polymer F-48 to remove the bulk of fibrinogen. The yield of Factor VIII at this point was 6356 theoretical and 42% actual (the starting plasma contained,66 units/ml VIII), Total protein in the cryoprecipitate was 2,2 gm# and 0.93 gn# after removal of fibrinogen bulk. The next step was addition of a thrombin-like enzyme, at a concentration of 0.5 units, with the resultant removal of remaining fibrinogen. After clotting out residual fibrinogen, resultant fibrin strands were removed by high speed centrifugation. The final product gave a theoretical yield of 65% and an actual yield of 28%; thus, 65% of the initial Factor VIII was recovered in the final product. Prior to lyophilization, the final product was stabilized with albumin. After lyophilization, no loss of activity occurred and solubility was excellent. Protein electrophoretic analysis revealed 70% in the albumin region, 7% in the alpha & beta globulin region, and 20% in the gamma region. Tests for hemolysins were negative and isoagglutin-in titers were 1:6. Plasminogen and plasmin were undetectable and fibrin(ogen) degradation products were 80 ug/ml. The large scale preparation of this fibrinogen-free Factor VIII concentrate may prove highly useful in sparing the hemophilic patient fibrin(ogen) deposits in the kidneys and other organs, a complication of existing concentrates. In addition, much higher potency in much less volume may be achieved with this material. The cost for large scale preparation of this material should be the same as for existing concentrates.

Sign in / Sign up

Export Citation Format

Share Document