Capturing biotinylated proteins and peptides by avidin functional affinity electrophoresis.
Avidin functional affinity electrophoresis (AFAEP) is a variational method of affinity electrophoresis. In this technique, avidin is immobilized within a small area of the gel matrix by interaction with acrylamide and/or polyacrylamide either directly or through bifunctional linker glutaraldehyde during polymerization. Analytes can be heated with Tris-glycine sodium dodecyl sulfate (SDS) sample buffer so that biotinylated peptides/proteins are negatively charged and migrate electrophoretically towards the cathode through the avidin zone regardless of their isoelectric point (pI) values. Alternatively, if the behavior of the biotinylated analytes is known, the SDS treatment is not required. The polarity of the electrodes is set such that biotinylated analytes migrate electrophoretically through the avidin zone. This technique can work with or without SDS in gel running buffer. The AFAEP method allows the capture and concentration of biotinylated peptides/proteins. The values of this technique stem from a combination of merits of polyacrylamide gel electrophoresis and affinity technology.
Lee
Bao-Shiang
BS
University of Chicago Research Resources Center, Chicago, IL, USA.
Krishnanchettiar
Sangeeth
S
Lateef
Syed Salman
SS
Gupta
Shalini
S
D019921:Biotinylation / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006023:Glycoproteins / Q000302:isolation & purification; D006801:Humans; D010455:Peptides / Q000302:isolation & purification*; D011506:Proteins / Q000302:isolation & purification*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
D006023:Glycoproteins; D010455:Peptides; D011506:Proteins