Quantification of target proteins using hydrogel antibody arrays and MALDI time-of-flight mass spectrometry (A2M2S).
Mass spectrometry-based analysis techniques are widely applied in proteomics. This study presents a novel method for quantitative multiplex candidate protein profiling. It applies immunocapture of differentially labeled protein complements on hydrogel antibody arrays and subsequent quantification by MS. To make this approach quantitative a labeling approach was devised. The impact of labeling on the antibody/antigen interaction was assessed in detail by surface plasmon resonance. Owing to there solution by mass more than two protein samples can be compared simultaneously. Direct labeling of crude samples such as sera was developed and so enables the absolute quantification of target proteins straight from crude samples without a protein purification step. It was used to measure the concentration of apolipoprotein A-1 in serum. This method has been termed A2M2S for Affinity Array sand MALDI Mass Spectrometry.
Darii
Ekaterina
E
CEA/Institut de Génomique, Centre National de Génotypage, Bâtiment G2, 2 rue Gaston Crémieux, 91057 Evry Cedex, France.
Lebeau
Diane
D
Papin
Nelly
N
Rubina
Alla Y
AY
Stomakhin
Andrei
A
Tost
Jörg
J
Sauer
Sascha
S
Savvateeva
Elena
E
Dementieva
Ekaterina
E
Zasedatelev
Alexander
A
Makarov
Alexander A
AA
Gut
Ivo G
IG
D020100:Hydrogels / Q000737:chemistry; D007118:Immunoassay / Q000379:methods*; D040081:Protein Array Analysis / Q000379:methods*; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D020100:Hydrogels; D011506:Proteins