Development of a Peptidomimetic Ligand for Efficient Isolation and Purification of Factor VIII via Affinity Chromatography

2007 ◽  
Vol 50 (18) ◽  
pp. 4329-4339 ◽  
Author(s):  
Sebastian Knör ◽  
Alexey V. Khrenov ◽  
Burkhardt Laufer ◽  
Evgueni L. Saenko ◽  
Charlotte A. E. Hauser ◽  
...  
Keyword(s):  
Factor Viii ◽  
Affinity Chromatography ◽  
Isolation And Purification

The Immunological Characterization of Human Antibodies to Factor VIII Isolated by Immuno-Affinity Chromatography

1981 ◽  
Vol 45 (01) ◽  
pp. 060-064 ◽  
Author(s):  
M L Kavanagh ◽  
C N Wood ◽  
J F Davidson
Keyword(s):  
Factor Viii ◽  
Affinity Chromatography ◽  
Gel Filtration ◽  
Immunological Characterization ◽  
Human Antibodies ◽  
Heavy Chains ◽  
Light Chain Type ◽  
Antibody Preparations ◽  
Chain Type

SummaryNine human antibodies to factor VIII were isolated from haemophilic plasmas by affinity chromatography and gel filtration and six were subsequently subjected to immunological characterization. Three partially purified preparations were similarly characterized. Eight of the antibodies were characterized as being exclusively IgG and one preparation was found to contain IgM. Seven of the antibodies contained only a single light chain type, four being of type lambda and three of type kappa. Two antibody preparations contained both kappa and lambda light chains. In four of the preparations, only a single heavy chain sub-class could be demonstrated, three of IgG3 and one of IgG4. Of the remainder, three were a mixture of IgG3 and IgG4 sub-classes and one contained both IgG2 and IgG4. IgG sub-classification could not be achieved with the IgM-containing preparation. These results demonstrate a restricted heterogeneity of light and heavy chains in human antibodies to factor VIII.

Peptide affinity chromatography of human clotting factor VIII

1998 ◽  
Vol 715 (1) ◽  
pp. 191-201 ◽  
Author(s):  
Roman Necina ◽  
Karin Amatschek ◽  
Eva Schallaun ◽  
Horst Schwinn ◽  
Djuro Josic ◽  
...  
Keyword(s):  
Factor Viii ◽  
Affinity Chromatography ◽  
Clotting Factor ◽  
Peptide Affinity ◽  
Human Clotting Factor

Development and validation of an affinity chromatography step using a peptide ligand for cGMP production of factor VIII

2004 ◽  
Vol 87 (3) ◽  
pp. 400-412 ◽  
Author(s):  
Brian D. Kelley ◽  
Molly Tannatt ◽  
Robert Magnusson ◽  
Sigrid Hagelberg ◽  
James Booth
Keyword(s):  
Factor Viii ◽  
Affinity Chromatography ◽  
Peptide Ligand ◽  
Cgmp Production ◽  
Development And Validation ◽  
Chromatography Step

Ferrochelatase: Isolation and purification via affinity chromatography

1980 ◽  
Vol 96 (2) ◽  
pp. 777-784 ◽  
Author(s):  
K. Mailer ◽  
R. Poulson ◽  
D. Dolphin ◽  
A.D. Hamilton
Keyword(s):  
Affinity Chromatography ◽  
Isolation And Purification

scholarly journals Isolation and purification of chloroplastic spinach (Spinacia oleracea) sedoheptulose-1,7-bisphosphatase

1987 ◽  
Vol 241 (1) ◽  
pp. 71-74 ◽  
Author(s):  
F Cadet ◽  
J C Meunier ◽  
N Ferté
Keyword(s):  
Affinity Chromatography ◽  
Spinacia Oleracea ◽  
Higher Plant ◽  
Isolation And Purification ◽  
Fructose 1,6 Bisphosphate ◽  
Molecular Sieving

Higher-plant sedoheptulose-1,7-bisphosphatase was isolated and purified over 200-fold from spinach (Spinacia oleracea) chloroplast stromal extracts to apparent electrophoretic homogeneity by DEAE-Fractogel, molecular sieving on Sephadex G-200 and Blue B dye-matrix affinity chromatography. It is a protein of Mr 66,000, made up of two apparently identical subunits (Mr 35,000). The enzyme is activated by reduced thioredoxin fb in the presence of dithiothreitol. Its specificity towards sedoheptulose 1,7-bisphosphate versus fructose 1,6-bisphosphate is high, but not absolute.

Affinity Chromatography of Human Factor VIII Using Human and Rabbit Antibodies to Factor VIII

1979 ◽  
Vol 42 (04) ◽  
pp. 1306-1315 ◽  
Author(s):  
Janet L Lane ◽  
H Ekert ◽  
A Vafiadis
Keyword(s):  
Molecular Weight ◽  
Factor Viii ◽  
Affinity Chromatography ◽  
Protein Concentration ◽  
Gel Filtration ◽  
Subunit Structure ◽  
Molecular Weights ◽  
Human Factor Viii ◽  
Sds Page ◽  
Normal Human

SummaryFactor VIII, purified by gel filtration on Sepharose 2B, has an 8 band multiple subunit structure, with molecular weights ranging from 30,000 to 230,000, on reduction and SDS-PAGE at a protein concentration of 400 μg/gel. Affinity chromatography of this factor VIII preparation with insolubilized haemophilic antibody to factor VIII showed that 45-81% VIII:C and 0-33% VIILRag were attached to the column. Elution of the column with 0.25 M CaCl2 did not show VIII:C or VIILRag in the eluate. NH4SCN dissociation of the column, followed by reduction and SDS-PAGE of the dissociated protein, showed that 95 % of the protein bound by haemophilic antibody had a molecular weight similar to the low molecular weight subunits of the reduced factor VIII.In control experiments with normal Human IgG, 3% of VIII:C and 5% of VIILRag were attached to the column. NH4SCN dissociation of the column, followed by reduction and SDS-PAGE of the protein, showed 2 faint bands with molecular weight consistent with heavy and light chains of IgG.Similar experiments with antibody to factor VIII showed that 67-83% of VIILC and 61-76% of VIII:Rag were attached to the column. Elution of the column with 0.25 M CaCl2 showed 10% of the applied VIII:C, but no VIII:Rag in the eluate. NH4SCN dissociation of the column, followed by reduction and SDS-PAGE of the dissociated protein, showed an 8 band subunit structure similar to the reduced factor VIII.

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