Characterization of Adenovirus Structural Proteins for Gene Therapy

6 Characterization of Adenovirus Structural Proteins for Gene Therapy 

LCYMS ANALYSIS OF PROTEINS AND PEPTIDES IN DRUG DISCOVERY 

For SDS-PAGE, adenovirus structural proteins dissociated from 1.0 x 10 particles/mL of virus were loaded onto each well of Novex precast gradient 4% to 20% acrylamide gels and stained with Coomassie Blue. 

Preparative RP-HPLC separation of viral proteins was performed on a 4.6-X 250-mm Jupiter C4 column at 50°C. Approximately 6mL of the column-purihed virus preparation ( 1 x 10 particles/mL) was loaded onto the column. Each chromatographic peak was collected manually for further MS characterization. 

The proteins separated by SDS-PAGE with MWs ranging from 10,000 Da to 100,000 Da were identified as adenoviral proteins from the SwissProt database when searched against all taxonomy. The results were summarized in Table 19-7. 

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