PMID- 28277648
OWN - NLM
STAT- MEDLINE
DCOM- 20190118
LR  - 20190118
IS  - 1520-6882 (Electronic)
IS  - 0003-2700 (Linking)
VI  - 89
IP  - 6
DP  - 2017 Mar 21
TI  - Sample Stacking Provides Three Orders of Magnitude Sensitivity Enhancement in SDS 
      Capillary Gel Electrophoresis of Adeno-Associated Virus Capsid Proteins.
PG  - 3285-3292
LID - 10.1021/acs.analchem.6b02933 [doi]
AB  - Size-based protein analysis utilizing only 25 ng of total proteins has been 
      realized by sodium dodecyl sulfate capillary gel electrophoresis (SDS CGE) with 
      head-column field-amplified sample stacking as an online sample preconcentration 
      technique. This method has been used as a replacement of SDS-PAGE for purity 
      analysis of adeno-associated virus (AAV) therapeutic products of different 
      serotypes and transgenes. A limit of detection of 0.2 ng/mL (3.3 pM) capsid 
      proteins was achieved with convenient UV absorbance detection at 214 nm, 
      equivalent to 20 pg of protein (330 attomole) loaded in the autosampler vial. For 
      purity analysis, only 25 ng of total AAV capsid proteins (4.3 femtomole virus 
      particles) were loaded to the autosampler vial. The sensitivity is comparable to 
      silver-stained SDS-PAGE. The RSD of purity measurement was 0.0-0.8%, comparable 
      to conventional SDS CGE utilizing 0.1-0.5 mg proteins. The new method provided 3 
      orders of magnitude sensitivity enhancement as compared to conventional SDS CGE. 
      It shares all the advantages of conventional SDS CGE (labor-saving, easy 
      automation, and convenient quantitation) and also the high sensitivity of silver 
      stained SDS-PAGE. The sample stacking SDS CGE technique can be adopted for 
      size-based analysis of other types of proteins. It is especially useful when 
      protein quantity or concentration is not sufficient for regular SDS CGE or 
      SDS-PAGE assay.
FAU - Zhang, Chao-Xuan
AU  - Zhang CX
AUID- ORCID: 0000-0003-1781-1582
AD  - Department of Therapeutics Production and Quality, St. Jude Children's Research 
      Hospital , Memphis, Tennessee 38105, United States.
FAU - Meagher, Michael M
AU  - Meagher MM
AD  - Department of Therapeutics Production and Quality, St. Jude Children's Research 
      Hospital , Memphis, Tennessee 38105, United States.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20170309
PL  - United States
TA  - Anal Chem
JT  - Analytical chemistry
JID - 0370536
RN  - 0 (Capsid Proteins)
RN  - 368GB5141J (Sodium Dodecyl Sulfate)
SB  - IM
MH  - Capsid Proteins/*analysis/chemistry
MH  - Dependovirus/*chemistry
MH  - Electrophoresis, Capillary/*methods
MH  - Sodium Dodecyl Sulfate/*chemistry
EDAT- 2017/03/10 06:00
MHDA- 2019/01/19 06:00
CRDT- 2017/03/10 06:00
PHST- 2017/03/10 06:00 [pubmed]
PHST- 2019/01/19 06:00 [medline]
PHST- 2017/03/10 06:00 [entrez]
AID - 10.1021/acs.analchem.6b02933 [doi]
PST - ppublish
SO  - Anal Chem. 2017 Mar 21;89(6):3285-3292. doi: 10.1021/acs.analchem.6b02933. Epub 
      2017 Mar 9.