scholarly journals AAV Production Everywhere: A Simple, Fast, and Reliable Protocol for In‐house AAV Vector Production Based on Chloroform Extraction

2020 ◽  
Vol 93 (1) ◽  
Author(s):  
Matilde Negrini ◽  
Gang Wang ◽  
Andreas Heuer ◽  
Tomas Björklund ◽  
Marcus Davidsson
Keyword(s):  
Aav Vector ◽  
Chloroform Extraction

Barcoded Rational AAV Vector Evolution Enables Systematic In Vivo Mapping of Peptide Binding Motifs

2018 ◽  
Author(s):  
Marcus Davidsson ◽  
Gang Wang ◽  
Patrick Aldrin-Kirk ◽  
Tiago Cardoso ◽  
Sara Nolbrant ◽  
...  
Keyword(s):  
Peptide Binding ◽  
Aav Vector ◽  
Binding Motifs

Application of RT-PCR for the Detection of Enteric Viruses in Oysters

1993 ◽  
Vol 27 (3-4) ◽  
pp. 49-53 ◽  
Author(s):  
L. A. Jaykus ◽  
R. De Leon ◽  
M. D. Sobsey
Keyword(s):  
Enteric Viruses ◽  
Rt Pcr ◽  
Chloroform Extraction ◽  
Cationic Detergent

Oyster samples processed by adsorption-elution-precipitation were seeded with poliovirus 1 and HAV, and cleaned and concentrated by Freon extraction (2X), PEG precipitation and chloroform extraction. Freon extraction resulted in recoveries of 63-76% for polio and 42-52% for HAV. PEG precipitation/chloroform extraction gave recoveries of 47-50% for polio and 15-19% for HAV. Treated extracts inhibited RT-PCR at 10−2 dilutions. Inhibitors were removed by treatment with the cationic detergent CTAB or Pro-Cipitate/UF adsorption-elution-concentration. Both treatments resulted in samples on which direct RT-PCR was possible. The CTAB procedure was able to detect 78 pfu of polio and 295 pfu of HAV. The Pro-Cipitate procedure was able to detect 70 pfu polio and 2.1×103 pfu HAV.

scholarly journals Alginate hydrogel polymers enable efficient delivery of a vascular-targeted AAV vector into aortic tissue

2021 ◽  
Author(s):  
Anca Remes ◽  
Dima Ibrahim Basha ◽  
Thomas Puehler ◽  
Christopher Borowski ◽  
Susanne Hille ◽  
...  
Keyword(s):  
Aortic Tissue ◽  
Alginate Hydrogel ◽  
Aav Vector ◽  
Efficient Delivery ◽  
Hydrogel Polymers

scholarly journals DETERMINATION OF FREE AND TOTAL CHOLESTEROL BY DIRECT CHLOROFORM EXTRACTION

1949 ◽  
Vol 180 (1) ◽  
pp. 315-328 ◽  
Author(s):  
George R. Kingsley ◽  
Roscoe R. Schaffert
Keyword(s):  
Total Cholesterol ◽  
Chloroform Extraction

scholarly journals How long is long-term? Delivery of anti-HIV antibodies using AAV vector

2019 ◽  
Vol 5 ◽  
pp. 49
Author(s):  
J. Martinez-Navio ◽  
R. Desrosiers ◽  
S. Fuchs ◽  
D. Mendes ◽  
E. Rakasz ◽  
...  
Keyword(s):  
Aav Vector ◽  
Term Delivery ◽  
Hiv Antibodies ◽  
Anti Hiv

Posttranslational modifications of recombinant myotube-synthesized human factor IX

Blood ◽  
2001 ◽  
Vol 97 (1) ◽  
pp. 130-138 ◽  
Author(s):  
Valder R. Arruda ◽  
James N. Hagstrom ◽  
Jeffrey Deitch ◽  
Terry Heiman-Patterson ◽  
Rodney M. Camire ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Posttranslational Modifications ◽  
Half Life ◽  
Specific Activity ◽  
Factor Ix ◽  
Serine Phosphorylation ◽  
Hemophilia B ◽  
Tyrosine Sulfation ◽  
Aav Vector ◽  
Carbohydrate Analysis

Abstract Recent data demonstrate that the introduction into skeletal muscle of an adeno-associated viral (AAV) vector expressing blood coagulation factor IX (F.IX) can result in long-term expression of the transgene product and amelioration of the bleeding diathesis in animals with hemophilia B. These data suggest that biologically active F.IX can be synthesized in skeletal muscle. Factor IX undergoes extensive posttranslational modifications in the liver, the normal site of synthesis. In addition to affecting specific activity, these posttranslational modifications can also affect recovery, half-life in the circulation, and the immunogenicity of the protein. Before initiating a human trial of an AAV-mediated, muscle-directed approach for treating hemophilia B, a detailed biochemical analysis of F.IX synthesized in skeletal muscle was carried out. As a model system, human myotubes transduced with an AAV vector expressing F.IX was used. F.IX was purified from conditioned medium using a novel strategy designed to purify material representative of all species of rF.IX in the medium. Purified F.IX was analyzed by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), N-terminal sequence analysis, chemical γ-carboxyglutamyl analysis, carbohydrate analysis, assays for tyrosine sulfation, and serine phosphorylation, and for specific activity. Results show that myotube-synthesized F.IX has specific activity similar to that of liver-synthesized F.IX. Posttranslational modifications critical for specific activity, including removal of the signal sequence and propeptide, and γ-carboxylation of the N-terminal glutamic acid residues, are also similar, but carbohydrate analysis and assessment of tyrosine sulfation and serine phosphorylation disclose differences. In vivo experiments in mice showed that these differences affect recovery but not half-life of muscle-synthesized F.IX.

Sign in / Sign up

Export Citation Format

Share Document